| human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain. | the adenovirus fiber protein is responsible for attachment of the virion to cell surface receptors. the identity of the cellular receptor which mediates binding is unknown, although there is evidence suggesting that two distinct adenovirus receptors interact with the group c (adenovirus type 5 [ad5]) and the group b (ad3) adenoviruses. in order to define the determinants of adenovirus receptor specificity, we have carried out a series of competition binding experiments using recombinant native f ... | 1995 | 7707507 |
| the n-terminal region of the adenovirus type 5 e1a proteins can repress expression of cellular genes via two distinct but overlapping domains. | the transforming e1a 12s and e1a 13s proteins of human adenovirus type 5 (ad5) contain two and three conserved regions, respectively. in the present study, the contribution of sequences in the nonconserved n-terminal region of the e1a proteins to morphological transformation and to down-regulation of a number of mitogen-inducible genes was investigated. as described previously, transformation of nrk cells (an established normal rat kidney cell line) results in denser cell growth and a cuboidal c ... | 1995 | 7707522 |
| replication-deficient adenovirus induces expression of interleukin-8 by airway epithelial cells in vitro. | preclinical studies with first-generation adenovirus (ad) vectors administered in vivo to the respiratory tract have demonstrated a nonspecific host response consisting, in part, of parenchymal neutrophil accumulation followed by mononuclear cell and macrophage accumulation. we hypothesized that the mechanism for this host response might be the elaboration of interleukin-8 (il-8) and monocyte chemoattractant protein-1 (mcp-1) from the airway epithelium following the exposure to ad. to evaluate t ... | 1995 | 7734515 |
| induction by adenovirus-5 e1a of the differentiation phenotype of f9 teratocarcinoma cells involves a conserved region (cr1) of e1a. | the effects of the e1a protein of adenovirus-5 on the differentiation program of f9 teratocarcinoma cells were examined by the stable introduction of plasmids that expressed wild-type or mutated forms of e1a. constitutive expression of plasmids for most of the mutant e1as induced loss of expression of the cell-surface antigen ssea-1 and the enhanced expression of genes specific for the differentiated phenotype of f9 cells, such as genes for laminin b1, tissue-type plasminogen activator (tpa) and ... | 1995 | 7742380 |
| activation of expression of multiple subfamilies of human alu elements by adenovirus type 5 and herpes simplex virus type 1. | the nearly one million alu repetitive elements in the human genome can be grouped into a number of subfamilies. comparisons between subfamily consensus sequences suggest that alu evolution is characterized by the sequential amplification and dispersal of a limited number of alu founder sequences. the s, sb and sb1 subfamilies provide an example of such a related series of alu subfamilies. we have previously demonstrated that adenovirus type 5 and herpes simplex virus type 1 activate rna polymera ... | 1995 | 7752221 |
| her-2/neu-targeting cancer therapy via adenovirus-mediated e1a delivery in an animal model. | overexpression of her-2/neu has been demonstrated in human ovarian cancer and correlated with poor prognosis. we previously found that the adenovirus type 5 early region 1a (e1a) gene product can repress overexpression and suppress the tumorigenic potential of the her-2/neu-overexpressing cancer cells. to develop an efficient her-2/neu-targeting gene therapy with e1a, a replication-deficient adenovirus containing the e1a gene, ad.e1a(+), was used to transduce the her-2/neu-overexpressing human o ... | 1995 | 7761095 |
| immunodetection of adenoviral e1a proteins in human lung tissue. | the adenoviral e1a proteins possess the ability to associate with the dna binding domains of a number of transcription factors, and in this manner promiscuously to activate a wide variety of genes. the present study was designed to determine whether this protein is expressed in human lungs where nonlytic adenoviral infection has been demonstrated. lung tissue from 12 patients harboring trace amounts of viral dna were examined along with a549 cells infected with adenovirus 5 and uninfected graham ... | 1995 | 7766428 |
| adenovirus type 5 precursor terminal protein-expressing 293 and hela cell lines. | hela and 293 cell lines that express biologically active adenovirus type 5 precursor terminal protein (ptp) have been made. the amount of ptp synthesized in these cell lines ranges from barely detectable to greater than that observed in cells infected with the wild-type virus. the ptp-expressing cell lines permit the growth of a temperature-sensitive terminal protein mutant virus sub100r at the nonpermissive temperature. a higher percentage of the stably transfected 293 cell lines expressed term ... | 1995 | 7769665 |
| foreign gene expression by human adenovirus type 5-based vectors studied using firefly luciferase and bacterial beta-galactosidase genes as reporters. | adenovirus (ad) vectors have been used extensively to obtain high-level expression of foreign genes in mammalian cells and are currently being studied for use as live viral-vectored vaccines and as gene transfer vectors for gene therapy. many ad recombinants have been generated that express foreign genes inserted in early region 3 (e3); however, little has been done to study the importance for gene expression of regulatory sequences flanking the gene. we have generated a series of ad5 helper-ind ... | 1995 | 7793076 |
| latent adenoviral infection in follicular bronchiectasis. | the follicular form of bronchiectasis originally described by whitwell (1) has been associated with adenoviral infection. the present study compares resected lungs from 16 patients with follicular bronchiectasis (in 10 of whom a nonviral etiology was identified) with those from eight patients with a nonfollicular histologic pattern. dna isolated from sections of 45 paraffin-embedded lung samples was subjected to the polymerase chain reaction (pcr) using primers for the e1a region of the adenovir ... | 1995 | 7812549 |
| mitotic cycle reactivation in terminally differentiated cells by adenovirus infection. | different cell types (e.g., neurons, skeletal and heart myocytes, adipocytes, keratinocytes) undergo terminal differentiation, in which acquisition of specialized functions entails definitive withdrawal from the cell cycle. such cells are distinct from quiescent (reversibly growth-arrested) cells, such as contact-inhibited fibroblasts. terminally differentiated cells can not be induced to proliferate by means of growth factor stimulation or transduction of cellular oncogenes. an important first ... | 1995 | 7814449 |
| a p53 mutation in exon 5 associated with adenovirus transformation. | the adenovirus type 5 (ad5) 55-kda e1b oncoprotein has been shown to form complexes with the p53 tumor suppressor protein. these complexes are thought to interfere with normal p53 activity and may be responsible for the paucity of p53 mutations in cells transformed by these viruses. this report describes an example of a p53 mutation in exon 5 in an ad5-transformed cell line that exhibited less expression of e1b 55-kda protein and a longer tumor-latency phenotype than another ad5-transformed cell ... | 1995 | 7818760 |
| the metabolism of small cellular rna species during productive subgroup c adenovirus infection. | during the late phase of subgroup c adenovirus infection, export of cellular mrna from the nucleus to the cytoplasm is inhibited. in one approach to investigate the mechanism whereby viral late mrnas are selected for export, we have examined the metabolism of small cellular rna species transcribed by all three rna polymerases during the late phase of ad5 infection. no changes in the quantities of [3h]uridine-labeled 5s rrna or trnas entering the cytoplasm were observed in infected cells. adenovi ... | 1995 | 7831765 |
| hpmpc, a broad-spectrum topical antiviral agent, inhibits herpes simplex virus type 1 replication and promotes healing of dendritic keratitis in the new zealand rabbit ocular model. | previously, we demonstrated that hpmpc, a new, broad-spectrum antiviral agent, inhibited adenovirus type 5 in the new zealand (nz) rabbit ocular model (cornea 1992; 11:529-33). historically, no antiviral agent has been demonstrated to be effective against both herpes simplex virus type 1 (hsv-1) and adenovirus eye infections in an experimental animal model. in this study, we compared topical 0.2% hpmpc to 1% trifluridine and vehicle control in the nz rabbit hsv-1 keratitis model. using a double- ... | 1994 | 7842711 |
| use of the colonic carcinoma cell line caco-2 for in vivo amplification and detection of enteric viruses. | the use of the continuous cell line caco-2 as an in vivo amplification system for the detection of fastidious human enteric viruses is reported. caco-2 cells showed an increased sensitivity to laboratory strains of group a rotavirus 3, reovirus 3, astrovirus 1, poliovirus 1, coxsackievirus a 24, enterovirus 70, and adenovirus 5, 40 and 41, when compared to a routine host cell line for each virus. nucleic acids from wild-type infectious rotavirus, astrovirus, and adenovirus 40 in stool samples of ... | 1994 | 7852976 |
| a permanent glial precursor cell line, immortalized with the adenovirus e1a gene, undergoes apoptosis in restrictive growth conditions. | we have previously described some differentiation properties of the erd.1.1 cell line, obtained after transfer and integration of the adenovirus-5 e1a gene. depending on the growth conditions, these cells differentiate towards the astrocyte or early oligodendrocyte differentiation pathway. however, in growth restrictive conditions, we observed dying cells that detached from the monolayer constituted of differentiating cells. this led us to examine the characteristics of the dying cells. the stud ... | 1995 | 7864853 |
| endotoxins and prednisolone alter replication of type 5 adenovirus and its temperature sensitive mutants. | latency, replication or transformation by adenoviruses require cooperation between their gene products and cellular factors, which are controlled by external stimuli. clinical observations suggest that bacterial endotoxin (lps) and steroid hormones have direct effects on the viral permissivity and activation. therefore, hep-2 cultures were infected with low multiplicity of wild type (wt) human adenovirus 5 (ad-5) and its temperature-sensitive mutants ts18 and ts19 damaged in the phosphorylation ... | 1994 | 7866727 |
| long-term episomal gene delivery in human lymphoid cells using human and avian adenoviral-assisted transfection. | human b-lymphoblastoid cells transformed by epstein-barr virus (ebv), a common source of established human cell lines, are generally refractory to standard dna transfection methods. we report here the use of human and chicken adenoviruses for the stable delivery of plasmid dna carrying the latent origin of replication, orip, from ebv into human b-lymphoblastoid cells. long-term hygromycin-resistant transformants of human lymphoblastoid cells belonging to the genetically unstable inherited diseas ... | 1994 | 7873181 |
| regulation of human retroviral latency by the nf-kappa b/i kappa b family: inhibition of human immunodeficiency virus replication by i kappa b through a rev-dependent mechanism. | the cellular transcription factor nf-kappa b stimulates human immunodeficiency virus type 1 (hiv-1) transcriptional initiation, but its role in the retroviral life cycle has not been fully defined. in this report, we show that i kappa b alpha acts as a cellular inhibitor of human retroviral replication through a discrete mechanism, independent of its effect on hiv transcription. i kappa b alpha inhibited hiv replication and gp160 expression by negatively regulating rev function, most likely acti ... | 1995 | 7878004 |
| vaccination of pigs with replication-defective adenovirus vectored vaccines: the example of pseudorabies. | the efficacy of a recombinant human adenovirus type 5 expressing gd, one of the immunogenic glycoprotein of pseudorabies virus, was tested in pigs. due to the deletion of the e1a gene, the recombinant virus is unable to replicate in non transcomplementing cells but is capable of eliciting an immune response against gp50 after inoculation into animals. the virus was formulated in a water/oil/water emulsion, a strategy previously shown to enhance the immune response against the virus-induced gp50. ... | 1994 | 7886933 |
| reexpression of neu-encoded oncoprotein counteracts the tumor-suppressing but not the metastasis-suppressing function of e1a. | by transfecting the adenovirus 5 e1a gene into neu-transformed nih3t3 cells, we previously showed that e1a can dramatically repress neu-encoded p185 expression and, concomitantly, suppress the features of transformation and metastasis of neu+e1a transfectants. from these results we concluded that suppression of transformation and metastasis by e1a in neu-transformed cells may be through repression of neu gene expression. however, e1a has recently been shown to also repress the transformation fea ... | 1993 | 7902209 |
| negative regulation of the neu promoter by the sv40 large t antigen. | the neu gene is amplified and its protein product is overexpressed in certain human tumors. the adenovirus 5 e1a gene product and c-myc repress neu transcription. moreover, expression of e1a in neu-transformed cells leads to decrease in transformation phenotype and metastatic potential. the simian virus 40 large t antigen (lt) shares structural and functional homology with e1a and c-myc, and all three proteins bind to the retinoblastoma gene product, rb. we found that lt also represses neu expre ... | 1993 | 7906953 |
| several hydrophobic amino acids in the p53 amino-terminal domain are required for transcriptional activation, binding to mdm-2 and the adenovirus 5 e1b 55-kd protein. | the p53 tumor suppressor gene product is a transcriptional activator that may be associated with its ability to suppress tumor cell growth. the acidic amino terminus of the p53 protein has been shown to contain this trans-activation activity as well as the domains for mdm-2 and adenovirus 5 e1b 55-kd protein binding. an extensive genetic analysis of this amino-terminal p53 domain has been undertaken using site-specific mutagenesis. the results demonstrate that the acidic residues in the amino te ... | 1994 | 7926727 |
| effect of cd4 gene expression on adenovirus replication. | the gene encoding the cd4 receptor was introduced into kb cells to establish the kbt4 cell line, a cell line susceptible to infection with human immunodeficiency virus type 1. adenovirus replication was found to be significantly less in these cells than in the parental kb cells. similar decreased adenovirus type 5 (ad5) replication occurred in helat4 cells compared with the original hela cells. the presence of cd4 did not alter the cell surface population of kb cell adenovirus receptors, since v ... | 1994 | 7933112 |
| early region 3-replacement adenovirus recombinants are less pathogenic in cotton rats and mice than early region 3-deleted viruses. | adenovirus type-5 (ad5) recombinant viruses with replacement of the 1.9 kb xbai fragment in the early region 3 (e3) by foreign genes have been constructed with the ultimate goal of inducing immune responses to the product of the inserted gene against a variety of virus infections. the pathogenicity of these recombinants, however, has not been studied. | 1994 | 7933985 |
| multiple pathways for activation of e2a expression in human kb cells by the 243r e1a protein of adenovirus type 5. | adenovirus type 5 (ad5) mutant dl520, which produces only the smaller 243 residue (243r) e1a protein, induced efficient production of the viral e2a 72-kda dna binding protein (dbp) in human kb cells, but not in human wi38, 143, or hela cells. in transient expression assays, the 243r e1a protein induced transcription from the e2 early promoter in kb but not in hela cells; there was no transcription from the e3 promoter in either cell line. in kb cells, truncation of the e2 promoter from -285 to - ... | 1994 | 7941702 |
| in vivo evaluation of the safety of adenovirus-mediated transfer of the human cystic fibrosis transmembrane conductance regulator cdna to the lung. | cystic fibrosis (cf) is a common, fatal hereditary disease resulting from mutations of the human cystic fibrosis transmembrane conductance regulator (cftr) gene in which epithelial cells throughout the body manifest altered regulation of apical membrane chloride secretion. although the disease affects multiple organs throughout the body, over 90% of patients die of complications of the lung involvement. the feasibility of adenovirus-derived vectors for in vivo delivery of the human cftr cdna to ... | 1994 | 7948135 |
| stimulation of erythropoiesis by in vivo gene therapy: physiologic consequences of transfer of the human erythropoietin gene to experimental animals using an adenovirus vector. | erythropoietin (epo), a 30.4-kd glycoprotein, is the principal regulator of erythropoiesis. to evaluate the concept that in vivo gene transfer might be used as an alternative to recombinant human epo (rhepo) in applications requiring a 1- to 3-week stimulation of erythropoiesis, the replication-deficient recombinant adenovirus admlp.epo was constructed by deleting the majority of e1 from adenovirus type 5, and replacing e1 with an expression cassette containing the adenovirus type 5 major late p ... | 1994 | 7949166 |
| topical hpmpc inhibits adenovirus type 5 in the new zealand rabbit ocular replication model. | to evaluate the antiviral inhibitory activity of hpmpc against ocular adenoviral serotypes in vitro and to determine the therapeutic efficacy and ocular toxicity of treatment with topical hpmpc on established adenovirus type 5 (ad5) mcewen infection in the new zealand (nz) rabbit ocular replication model. | 1994 | 7960596 |
| characterisation of australian ovine adenovirus isolates. | we have characterised two groups of adenoviruses isolated from sheep in australia. restriction endonuclease maps for enzymes bamhi, clai, sali, smai and sphi have been determined for the genome of ovine adenoviruses related to bovine adenovirus serotype 7 (bav 7) from sheep in western australia. although previously serotyped as bav 7 these isolates are different from bovine isolates of bav 7 based on comparison with published restriction endonuclease profiles and maps of bav 7 cattle isolates. a ... | 1994 | 7975153 |
| a precursor terminal protein-trinucleotide intermediate during initiation of adenovirus dna replication: regeneration of molecular ends in vitro by a jumping back mechanism. | the adenovirus type 5 origin sequence starts with 3' gtagta. initiation of replication occurs by a protein priming mechanism in which the viral precursor terminal protein (ptp) is covalently linked to the first nucleotide of the nascent chain, a dcmp residue. this suggests that a ptp-dcmp (ptp-c) complex functions as an initiation intermediate. employing a reconstituted replication system and both synthetic oligonucleotides and the natural tp-dna as templates, we show that ptp-cat rather than pt ... | 1994 | 7988575 |
| activity of ganciclovir against human adenovirus type-5 infection in cell culture and cotton rat eyes. | the most common causes of acute viral infections of the eye for which there are no effective antiviral drugs are the adenoviruses. until recently, pathogenesis studies and antiviral drug testing for adenovirus-induced ocular disease were not practical because no animal model was available. however, new animal models for human adenovirus-induced ocular and respiratory infections have now made such studies possible. we assessed the in vitro and in vivo activity of ganciclovir against a genetically ... | 1994 | 7995068 |
| ablation of e2a in recombinant adenoviruses improves transgene persistence and decreases inflammatory response in mouse liver. | first-generation recombinant adenoviruses that lack e1 sequences have shown tremendous promise in animal and human models of gene therapy. important limitations of these vectors are that recombinant gene expression is transient and inflammation occurs at the site of gene transfer. our hypothesis for generating vectors with increased persistence is that present recombinant adenoviruses express viral proteins that stimulate cellular immune responses leading to destruction of the infected cells and ... | 1994 | 8016137 |
| structural study of the 5' end of a synthetic premessenger rna from adenovirus. evidence for a long-range exon-intron interaction. | in order to establish the structural features of the cis-elements involved in splicing and in its regulation, we have analyzed a synthetic premessenger rna, derived from the e3 transcription unit of adenovirus-2 and previously shown to be a good substrate for in vitro splicing. the transcript was probed by enzymatic and chemical methods and we present the structure in solution of the upstream exon and of the 5' part of the intron. this 417 nucleotide long fragment, which overlaps the exon-intron ... | 1994 | 8028005 |
| compartmentalization of cellular and viral dnas in adenovirus type 5 infection as revealed by ultrastructural in situ hybridization. | we describe the respective distributions of cellular dna and viral genomes in adenovirus type 5 (ad5)-infected hela cells by means of electron microscope in situ hybridization using biotinylated alu dna and ad5 dna probes in a post-embedding technique. when hybridization was performed on sections of formaldehyde-fixed material, alu elements were restricted to the cellular chromatin, irrespective of the stage of the infectious cycle, without any interpenetration within the virus-induced regions. ... | 1994 | 8032671 |
| complementary functions of e1a conserved region 1 cooperate with conserved region 3 to activate adenovirus serotype 5 early promoters. | the amino-terminal region of the adenovirus type 5 e1a protein including conserved regions (crs) 1 and 2 binds the 105-kda retinoblastoma protein and a second, 300-kda, cellular protein. we show that mutant viruses with deletions of cr1 which release the binding of either p105 or p300 still activate early promoters and infect cells productively. however, mutations which disrupt binding of both proteins disrupt early promoter activity and block the viral life cycle. ela cr3, which has an establis ... | 1994 | 8035489 |
| characterization of the knob domain of the adenovirus type 5 fiber protein expressed in escherichia coli. | the adenovirus fiber protein is used for attachment of the virus to a specific receptor on the cell surface. structurally, the protein consists of a long, thin shaft that protrudes from the vertex of the virus capsid and terminates in a globular domain termed the knob. to verify that the knob is the domain which interacts with the cellular receptor, we have cloned and expressed the knob from adenovirus type 5 together with a single repeat of the shaft in escherichia coli. the protein was purifie ... | 1994 | 8035520 |
| dp-1: a cell cycle-regulated and phosphorylated component of transcription factor drtf1/e2f which is functionally important for recognition by prb and the adenovirus e4 orf 6/7 protein. | the cellular transcription factor drtf1/e2f integrates cell cycle events with the transcription apparatus through its cyclical interactions with important regulators of cellular proliferation. two sequence-specific dna binding proteins, dp-1 and e2f-1, are components of drtf1/e2f which synergistically interact in a dp-1/e2f-1 heterodimer. here, we show that dp-1 is a very frequent, possibly universal, component of drtf1/e2f in 3t3 cells since it is present in all forms of the dna binding activit ... | 1994 | 8039504 |
| cloning of the rat tissue inhibitor of metalloproteinases type 2 (timp-2) gene: analysis of its expression in normal and transformed thyroid cells. | we have recently reported that the introduction of the e1a gene of the adenovirus 5 gene into the rat thyroid pc cl 3 epithelial cell line causes loss of the differentiated thyroid phenotype without the appearance of the typical markers of neoplastic transformation. it is well known that the e1a gene is able either to induce or to repress the transcription of various endogenous cellular genes. in order to characterize the genes involved in the transformation of pc cl 3 cells by the e1a gene, we ... | 1994 | 8050496 |
| late transcripts of adenovirus type 12 dna are not translated in hamster cells expressing the e1 region of adenovirus type 5. | hamster cells are completely nonpermissive for the replication of human adenovirus type 12 (ad12), whereas types 2 and 5 can replicate in hamster cells. the ad5-transformed hamster cell line bhk297-c131, which carries the left terminal 18.7% of the ad5 genome and expresses at least the viral e1a region, can somehow complement ad12 dna replication and the transcription of the late ad12 genes. since the interaction of ad12 with hamster cells must constitute a significant factor in the induction of ... | 1994 | 8057430 |
| high resolution localization of replicating viral genome in adenovirus-infected hela cells. | previous autoradiographical and in situ hybridization experiments have revealed that the replication of viral genomes in adenovirus type 5 infected hela cells induces changes in nuclear structure of which one of the more striking is the formation of distinctive replicative foci. the latter consist of a viral ssdna accumulation site and a surrounding fibrillogranular network. we have reexamined these structures and processes by a more direct and higher resolution approach, that is, incorporation ... | 1994 | 8082651 |
| adenoviral-mediated gene transfer of human surfactant protein b to respiratory epithelial cells. | human surfactant protein b (sp-b) is a 79-amino acid, phospholipid-associated polypeptide expressed by respiratory epithelial cells of the lung. sp-b is essential for lung function, enhancing the spreading and stability of surfactant phospholipids that serve to reduce surface tension at the alveolar air-liquid interface. congenital absence of sp-b results in neonatal respiratory failure and death. in the present work, we constructed a replication-deficient adenoviral vector, av1sp-b1, in which t ... | 1994 | 8086169 |
| enhanced c-erbb-2/neu expression in human ovarian cancer cells correlates with more severe malignancy that can be suppressed by e1a. | amplification or overexpression of c-erbb-2/neu protooncogene, or both, occur frequently in many different types of human cancers and have been shown to correlate with decreased survival in ovarian cancer patients. we have previously found that the ovarian carcinoma cell line sk-ov-3 overexpresses c-erbb-2/neu mrna. to further study the biological effect of c-erbb-2/neu overexpression in sk-ov-3 cells, we injected such cells i.p. into female nu/nu mice and found that this cell line forms extensi ... | 1993 | 8094034 |
| chicken adenovirus (celo virus) particles augment receptor-mediated dna delivery to mammalian cells and yield exceptional levels of stable transformants. | delivery of genes via receptor-mediated endocytosis is severely limited by the poor exit of endocytosed dna from the endosome. a large enhancement in delivery efficiency has been obtained by including human adenovirus particles in the delivery system. this enhancement is probably a function of the natural adenovirus entry mechanism, which must include passage through or disruption of the endosomal membrane. in an effort to identify safer virus particles useful in this application, we have tested ... | 1993 | 8099627 |
| structure, function, and chromosome mapping of the growth-suppressing human homologue of the murine gas1 gene. | we describe the isolation, growth-suppressing activity, and chromosomal localization of the human homologue of the murine growth-arrest-specific gene gas1. overexpression of h-gas1 is able to block cell proliferation in the a549 lung carcinoma and the t24 bladder carcinoma cell lines. no effect was observed when h-gas1 was introduced into the osteosarcoma cell line saos-2 and into the adenovirus-type-5 transformed cell line 293. this finding is related to our previous evidence that simian virus ... | 1994 | 8127893 |
| induction of apoptosis by adenovirus type 5 e1a in rat cells requires a proliferation block. | infection with ad5dl520eib-, an adenovirus producing only the 243 residue e1a protein and lacking the e1b region, caused apoptosis in normal rat kidney (nrk) cells as judged by the production of nucleosomal dna fragments. apoptosis occurred only when the cells were growth-inhibited by cell-cell contacts in confluent cultures or by serum starvation and not when they were actively growing. in uninfected cultures, apoptosis also occurred at confluency, but more slowly than after infection. studies ... | 1994 | 8134121 |
| trans-dominant interference of type 5 adenovirus e1a mutants in cell transformation. | two type 5 adenovirus (ad5) early region 1a (e1a) mutants, h5in104 and h5dl105, were impaired in viral replication and cell transformation. in addition, these mutants trans dominantly inhibited the frequency with which h5sub309, a phenotypically wild-type mutant, and h5dl520, a high-frequency transformation mutant, transformed cref cells. inhibition of transformation varied in proportion to the input ratio of mutant to coinfecting virus. it was found that h5in104, but not h5dl105, could not comp ... | 1994 | 8138997 |
| characterization of the 55k adenovirus type 5 e1b product and related proteins. | in addition to major proteins of 19k and 55k (176 and 496 residues, 176r and 496r, respectively), early region 1b (e1b) of human adenovirus type 5 (ad5) is predicted to encode at least three other polypeptides of 156r, 93r and 84r that share 79 amino-terminal residues with 496r. we have used a series of specific antipeptide sera to identify and partially characterize these proteins. 84r was produced in large amounts, 156r somewhat less, and 93r at very low levels. synthesis of 176r, 496r, as wel ... | 1994 | 8151295 |
| incorporation of adenovirus into a ligand-based dna carrier system results in retention of original receptor specificity and enhances targeted gene expression. | adenovirus type 5 was modified by coupling an asialoglycoprotein-polylysine conjugate to the virus by reactions that activate carbohydrate residues. wild-type virus modified in this manner had greatly decreased infectivity toward normally susceptible hela s3 (asialoglycoprotein receptor (-)) and sk hep1 (asialoglycoprotein receptor (-)) cells leaving 91 and 86% viable, respectively, after 48 h. however, with huh 7 (asialoglycoprotein receptor (+)) cells, modified virus retained its infectivity l ... | 1994 | 8157685 |
| role of respiratory viral infection in sids: detection of viral nucleic acid by in situ hybridization. | there is considerable evidence suggesting that respiratory viral infection is involved in the genesis of the sudden infant death syndrome (sids), with rates of about 20 per cent of sids victims compared to about 13 per cent of controls. since the techniques used previously are prone to under-reporting from autopsy material, non-isotopic in situ hybridization (nish) has been used to detect viral nucleic acid in lung in sids. forty-five sids cases (30 males) were examined (age range 3 weeks-14 mon ... | 1993 | 8158456 |
| the e1 sequence of bovine adenovirus type 3 and complementation of human adenovirus type 5 e1a function in bovine cells. | the bovine adenovirus type 3 (bav3) genome was sequenced from the left end to the hindiii site at 11%. this region comprises the entire e1 transcription unit including the open reading frames (orf) for proteins homologous to the e1a, e1b proteins and protein ix of human adenovirus type 5 (ad5). a portion of the bav3 e1a protein showed significant homology with conserved region 3 (cr3), the principal transactivation region of ad5 e1a. the bav3 e1a protein also contains a consensus sequence known ... | 1994 | 8178572 |
| concurrent mapping of an adenovirus 5/sv40 integration site and the u1 snrna cluster (rnu1) within 400 kb of the chromosome region 1p36.1. | previous reports from our group suggested the preferential integration of the viral construct ad5/sv40 at the short arm subtelomeric region of human chromosome 1. the present study narrows the region of viral integration to site 1p36.1 in a close cytogenetic overlap with the u1 snrna gene cluster (rnu1) within a distance necessarily smaller than 400 kb as suggested by the size of the yac in which the two markers were found to coexist. this finding supports the hypothesis that the chromosomal sit ... | 1994 | 8187549 |
| inhibition of type 5 adenovirus infectivity by periodate oxidation. | periodate oxidation of purified type 5 adenovirus (ad5) led to a mean loss of infectivity of 6.84 logs. there were no significant differences in adsorption and penetration between oxidized and mock-oxidized virus. however, after infection with oxidized virus, no synthesis of viral structural proteins could be detected and a 78.5% inhibition of viral dna synthesis was observed. labelling experiments performed by treating oxidized and mock-oxidized virus with tritiated sodium borohydride revealed ... | 1994 | 8198449 |
| recovery of viruses other than cytomegalovirus from bronchoalveolar lavage fluid. | to determine the yield and diagnostic significance of performing viral cultures on specimens obtained by bronchoalveolar lavage (bal) in immunocompromised patients. | 1994 | 8205876 |
| role of membrane anchor domain of bcl-2 in suppression of apoptosis caused by e1b-defective adenovirus. | bcl-2 is an integral membrane protein that functions as a suppressor of programmed cell death. it contains a cooh-terminal signal anchor sequence that is selective for import and insertion of bcl-2 into the mitochondrial outer membrane and, by a different mechanism, can also direct the protein to other membrane sites. deletion of the signal anchor sequence rendered bcl-2 cytosolic and impaired its ability to prevent apoptotic death of human kb cells infected with a mutant form of adenovirus type ... | 1994 | 8206964 |
| the utility of latex agglutination assays in the diagnosis of pediatric viral gastroenteritis. | to design a rapid and efficient protocol for processing pediatric stool specimens, the authors used 434 specimens to evaluate two commercial latex assays to detect rotavirus (meritec-rotavirus and rotalex) and one to detect adenovirus (adenolex). rotavirus latex assay results were compared with electron microscopic examination and adenovirus latex assay results with virus culture. ninety-two specimens (21%) were positive for rotavirus and 28 (6.5%) for adenovirus; 5 (1%) had both viruses. the se ... | 1994 | 8209862 |
| genetic analysis of carcinogen enhancement of type 5 adenovirus transformation of cloned fischer rat embryo fibroblast cells. | pretreatment of cref cells with methyl methanesulfonate (mms) before infection with the host-range cold-sensitive type 5 adenovirus (ad5) mutant h5hr1 results in a dose-dependent carcinogen enhancement of viral transformation (cet). the properties of cet observed with h5hr1, which include both an mms dose-dependent enhancement in the number of transformed foci and an increase in transformation frequency after correction for cell toxicity, are not observed in carcinogen-pretreated wild-type (wt) ... | 1993 | 8216734 |
| induction of gene expression by exon 2 of the major e1a proteins of adenovirus type 5. | we have constructed an adenovirus type 5 (ad5) e1a mutant, dl1119/520, that produces essentially only exon 2 of the major e1a proteins. in infected primary baby rat kidney cells, this mutant induced expression of the e1b 55-kda protein, and in infected human kb cells, it induced expression of this protein, the e2a 72-kda protein, and hexon. in kb cells, this mutant grew substantially better than ad5 dl312, which lacks e1a, and as well as ad5 dl520, an e1a mutant producing only the 243-residue pr ... | 1993 | 8230413 |
| superiority of in situ hybridization over immunolabeling for detecting dna on lowicryl sections: a study on adenovirus-infected cells. | we investigated the intranuclear distribution of viral genomes in adenovirus type 5-infected hela cells on ultra-thin sections of lowicryl k4m-embedded material by immunolabeling of dna and in situ hybridization with a viral dna probe. monoclonal ac-30-10, raised against double- and single-stranded dna (dsdna, ssdna), and hb2 antibodies, raised against only the dsdna, were used. both antibodies intensely labeled the condensed host chromatin and the virus-induced substructures containing inactive ... | 1993 | 8245412 |
| analysis of a viral integration event in a cg-rich region at the 1p36 human chromosomal site. | the preinsertion site of an adenovirus-5/simian virus 40 recombinant construct (ad5/sv40) has been cloned and sequenced. our data suggest that viral integration has occurred in a genomic region which has been the target of multiple events of alu element retropositions within a taa minisatellite. extensive homologies between the left viral end and the host cellular dna were also observed. the compositional similarity between adenoviridae and the region of viral integration is consistent with the ... | 1993 | 8276253 |
| human adenovirus type 5 recombinants expressing simian immunodeficiency virus macaque strain gag antigens. | the p55 gag gene of simian immunodeficiency virus macaque strain (sivmac) and the core p27 gag component linked to a synthetic aug codon have been cloned into adenovirus type 5 vectors to generate either viable e3-replacement or defective e1-replacement viruses. the viruses express the expected siv proteins in both human and, for the non-defective viruses, monkey cells. a considerable proportion of the p55 produced is exported from the infected cell. these viruses should prove useful both in stu ... | 1993 | 8277293 |
| phosphorylation at the carboxy terminus of the 55-kilodalton adenovirus type 5 e1b protein regulates transforming activity. | the 55-kda product of early region 1b (e1b) of human adenoviruses is required for viral replication and participates in cell transformation through complex formation with and inactivation of the cellular tumor suppressor p53. we have used both biochemical and genetic approaches to show that this 496-residue (496r) protein of adenovirus type 5 is phosphorylated at serine and threonine residues near the carboxy terminus within sequences characteristic of substrates of casein kinase ii. mutations w ... | 1994 | 8289381 |
| tumorigenicity of adenovirus-transformed rodent cells is influenced by at least two regions of adenovirus type 12 early region 1a. | chimeric adenovirus type 5 (ad5)/ad12 early region 1a (e1a) genes were used to transform primary baby rat kidney cells in cooperation with ad12 e1b, and the resulting cell lines were assayed for tumorigenicity in syngeneic rats. it was found that lines were nontumorigenic when transformed by hybrid e1a genes consisting of the amino-terminal 80 amino acids from ad12 including conserved region 1 (cr1), with the remaining portion from ad5. in contrast, cell lines transformed by hybrids containing a ... | 1994 | 8289391 |
| highly efficient gene transfer into adult ventricular myocytes by recombinant adenovirus. | molecular dissection of mechanisms that govern the differentiated cardiac phenotype has, for cogent technical reasons, largely been undertaken to date in neonatal ventricular myocytes. to circumvent expected limitations of other methods, the present study was initiated to determine whether replication-deficient adenovirus would enable efficient gene transfer to adult cardiac cells in culture. adult rat ventricular myocytes were infected, 24 h after plating, with adenovirus type 5 containing a cy ... | 1993 | 8326005 |
| the e1a products of oncogenic adenovirus serotype 12 include amino-terminally modified forms able to bind the retinoblastoma protein but not p300. | the cell growth-regulating properties of the adenovirus type 5 (ad5) e1a oncogene correlate closely with the binding of the e1a products to specific cellular proteins. these proteins include the products of the retinoblastoma tumor susceptibility gene and a 300-kda product, p300. prb binds to e1a sequences that are highly conserved among the e1a products of various serotypes, while p300 binding requires sequences in the e1a amino terminus, a region that is not highly conserved. to help evaluate ... | 1993 | 8331729 |
| large e1b proteins of adenovirus types 5 and 12 have different effects on p53 and distinct roles in cell transformation. | the formation of complexes between oncoproteins of dna tumor viruses and the cellular protein p53 is thought to result in inactivation of the growth suppressor function of p53. in cells transformed by nononcogenic human adenovirus type 5 (ad5), the 55-kda protein encoded by e1b forms a stable complex with p53 and sequesters it in the cytoplasm. however, the homologous 54-kda protein of highly oncogenic ad12 does not detectably associate with p53. yet in ad12-transformed cells, p53 is metabolical ... | 1993 | 8350396 |
| protocol of electron microscope in situ nucleic acid hybridization for the exclusive detection of double-stranded dna sequences in cells containing large amounts of homologous single-stranded dna and rna sequences: application to adenovirus type 5 infected hela cells. | in order to gain a further insight into the relationships of the complex process of replication of adenovirus genomes to the substructures which occur in the nuclei of adenovirus type 5 (ad5) infected hela cells, we have visualized directly, at the electron microscopic level, viral double-stranded dna (dsdna) in late infected nuclei by the use of a post-embedding in situ hybridization technique with a biotinylated specific dna probe. the procedure is based on the removal of single-stranded (ss) ... | 1993 | 8353305 |
| regulation of hiv-1 envelope protein synthesis by tat and rev in 293 cells. | a plasmid expression vector (b2) with the hiv-1 envelope sequence downstream of the adenovirus type 5 early region 3 promoter could direct the synthesis of envelope protein in the absence of rev when transfected into 293 cells. we investigated this further using pnl4.3 delta tr, and hiv-1 mutant which lacks the first exon of tat and rev and pnl4.3 delta r, an hiv-1 mutant with a premature termination codon in the second coding exon of rev. in cells transfected with pnl4.3 delta tr and a tat-expr ... | 1993 | 8356789 |
| packaging capacity and stability of human adenovirus type 5 vectors. | adenovirus vectors are extensively used for high-level expression of proteins in mammalian cells and are receiving increasing attention for their potential use as live recombinant vaccines and as transducing viruses for use in gene therapy. although it is commonly argued that one of the chief advantages of adenovirus vectors is their relative stability, this has not been thoroughly investigated. to examine the genetic stability of adenovirus type 5 vectors and in particular to examine the relati ... | 1993 | 8371349 |
| pathogenesis of adenovirus type 5 pneumonia in cotton rats (sigmodon hispidus). | cotton rats (sigmodon hispidus) were inoculated intranasally with 10(2.0) to 10(10.0) pfu of human adenovirus type 5. the virus replicated to a high titer in pulmonary tissues, with the peak titer being proportional to the input dose. the 50% lethal dose was 10(9.4) pfu. histopathologic changes were proportional to the infecting inoculum and included the infiltration of interstitial and intra-alveolar areas, moderate damage to bronchiolar epithelium, and cellular infiltration of peribronchiolar ... | 1993 | 8380066 |
| stabilization of the p53 tumor suppressor is induced by adenovirus 5 e1a and accompanies apoptosis. | oncogenic transformation by human adenoviruses requires early regions 1a and 1b (e1a and e1b) and provides a model of multistep carcinogenesis. this study shows that the metabolic stabilization of p53 observed in adenovirus 5 (ad5)-transformed cells can occur in untransformed cells expressing e1a alone. stabilized p53 was localized to the nucleus and was indistinguishable from wild-type p53 with respect to its interactions with hsc70, pab420, ad5 p55e1b, and sv40 large t antigen. moreover, bindi ... | 1993 | 8384579 |
| protective immunity to rotavirus-induced diarrhoea is passively transferred to newborn mice from naive dams vaccinated with a single dose of a recombinant adenovirus expressing rotavirus vp7sc. | vp7sc is a novel rotavirus antigen engineered for presentation at the cell surface. several recombinant viruses were constructed in which vp7sc was inserted into the e3 region of the human type 5 adenovirus (ad5) genome and expression and transport of the antigen was monitored in cultured 293 cells. the recombinant virus showing the greatest level of expression (ad5/7.4) was then used to determine whether antibodies to vp7sc could be induced in a nonhuman host. balb/c and cba/h mice were inocula ... | 1993 | 8384761 |
| wild-type adenovirus type 5 transforming genes function as transdominant suppressors of oncogenesis in mutant adenovirus type 5 transformed rat embryo fibroblast cells. | transformation of cloned rat embryo fibroblast (cref) cells with the host-range adenovirus type 5 (ad5) mutant, h5hr1, results in transformants with a fibroblastic morphology which displays a cold-sensitive transformation phenotype and oncogenic potential in both nude mice and syngeneic rats. in contrast, wild-type (wt) ad5 transformed cref cells are epithelioid in morphology, temperature independent for transformation, and nontumorigenic. the present studies were conducted to analyze the contri ... | 1993 | 8385576 |
| monitoring foreign gene expression by a human adenovirus-based vector using the firefly luciferase gene as a reporter. | we have constructed a helper-independent adenovirus type 5-luciferase recombinant (ad5-luc 3) containing the firefly luciferase gene flanked by simian virus 40 (sv40) regulatory sequences inserted in the early region 3 (e3) of the ad5 genome. expression of luciferase in cells infected with ad5-luc3 was relatively efficient. in hela cells approximately 20 micrograms luciferase per 10(6) cells was made by 36 h post-infection and a 62 kilo-dalton (kda) luciferase band was clearly visible in a [35s] ... | 1993 | 8388142 |
| [use of ultrastructural in situ hybridization for the intracellular detection of viral dna and rna]. | post-embedding in situ nucleic acid hybridization (ish) at the electron microscope level is at present the best tool for the ultrastructural detection of specific viral sequences in infected cells since sensitivity and specificity of the method are paralleled by preservation of ultrastructure. biotin-labelled double-stranded viral dna probes were applied at the surface of lowicryl k4m sections for localizing viral nucleic acids (all viral dna, only single-stranded viral dna, or viral rna) in her ... | 1993 | 8392162 |
| in vivo detection of fluorescent tumor-specific cytotoxic t cell clones. | cytotoxic t lymphocytes (ctl) can be very effective mediators of tumor-specific immunity in vivo. since little is known about the in vivo behaviour of cultured tumor-specific ctl, a fast and simple method has been developed utilizing a lipophilic carbocyanine, 1,1'-dioctadecyl 3,3,3',3'-tetramethylin-docarbocyanine perchlorate (dii), for the in vivo detection of tumor-specific ctl clones in (tumor-bearing) mice. the two ctl clones used in this study are directed against human papillomavirus type ... | 1993 | 8393053 |
| biological and biochemical activity of e7 genes of the cutaneous human papillomavirus type 5 and 8. | in contrast to the observed activity of the e7 genes of the genital high-risk human papillomavirus (hpv)16 and hpv18, e7s of the cutaneous high-risk hpv5 and hpv8 show no in vitro transforming activity in established rodent cells. we recently reported that the hpv8 e7 driven by the sv40 enhancer/promoter oncogenically transforms primary rat embryo fibroblast (ref) cells collaboratively with the ejras oncogene (jpn. j. cancer res., 82, 1340-1343, 1991). to study the functional differences between ... | 1993 | 8395681 |
| mouse hepatitis virus spike and nucleocapsid proteins expressed by adenovirus vectors protect mice against a lethal infection. | infection with the mouse hepatitis coronavirus (mhv) provides an excellent model for the study of viral diseases of the central nervous system and the gastrointestinal tract. with the ultimate aim of studying mucosal immunity to mhv we have cloned the genes encoding the structural proteins of mhv strain a59 (mhv-a59) into the e3 region of a human adenovirus type 5 vector. infection of hela cells with the resulting recombinant adenoviruses admhvs, admhvn and admhvm revealed the correct expression ... | 1993 | 8409930 |
| multiple pathways for gene activation in rodent cells by the smaller adenovirus 5 e1a protein and their relevance to growth and transformation. | by immunoprecipitating protein products from virus-infected baby rat kidney (brk) cells with specific antibodies, we found that the smaller, 243 residue (243r) e1a protein of human adenovirus 5 (ad5) activated expression of the virus genes for e1b 55k, e2a 72k, e3 19k, hexon, fibre and penton base and the cellular gene for pcna. the 243r protein also activated the e2a 72k gene in several rodent cell lines. in transient expression assays, this protein trans-activated the e2 early and major late p ... | 1993 | 8409938 |
| analysis of the adenovirus type 5 terminal protein precursor and dna polymerase by linker insertion mutagenesis. | a series of adenovirus type 5 precursor terminal protein (ptp) and dna polymerase (ad pol) genes with linker insertion mutations were separately introduced into the vaccinia virus genome under the control of a late vaccinia virus promoter. the recombinant viruses were used for overexpression of the mutant genes in hela cells. in total, 22 different mutant ptp and 10 different ad pol vaccinia virus recombinants were constructed, including some that expressed carboxyl-terminus-truncated forms of b ... | 1993 | 8416372 |
| correction of a deletion mutant by gene targeting with an adenovirus vector. | the usefulness of adenovirus type 5 as a vector for homologous recombination was examined in cho cells by using the adenine phosphoribosyltransferase (aprt) gene. infection of a hemizygous cho aprt- cell line containing a 3-bp deletion in exon 5 of the aprt gene with a recombinant adenovirus containing the wild-type gene resulted in restoration of the aprt+ phenotype at a frequency of 10(-5) to 10(-6) per infected cell. a relatively high frequency (approximately 6 to 20%) of the transductants ap ... | 1993 | 8423811 |
| the adenovirus e1a gene blocks the differentiation of a thyroid epithelial cell line, however the neoplastic phenotype is achieved only after cooperation with other oncogenes. | the pc cl 3 cell line is a well characterized epithelial thyroid cell line of fischer rat origin. this cell line has the peculiarity of retaining in vitro the typical markers of thyroid differentiation (i.e. thyroglobulin synthesis and secretion, iodide uptake and dependence on tsh for growth). the pc cl 3 cells have been transfected with the e1a gene of adenovirus 5. the e1a transfected cells, pc e1a, partially lost the dependency on tsh for growth and completely lost the ability to trap iodide ... | 1993 | 8426735 |
| the e1b 19-kilodalton protein is not essential for transformation of rodent cells in vitro by adenovirus type 5. | the newly constructed adenovirus type 5 mutant in1 carries a single at base pair insertion immediately after nucleotide position 1715 in the e1b gene sequence which destroys the proximal aug normally present in e1b messages and prevents production of intact e1b 19-kda protein in infected cells. we have used in1, variants of in1 containing mutant alleles of viral genes known to enhance transformation frequency, and adenovirus type 5 mutant dl337 (s. pilder, j. logan, and t. shenk, j. virol. 52:66 ... | 1993 | 8437231 |
| characterization of mutants within the gene for the adenovirus e3 14.7-kilodalton protein which prevents cytolysis by tumor necrosis factor. | the 14,700-da protein (14.7k protein) encoded by the e3 region of adenovirus has previously been shown to protect mouse cells from cytolysis by tumor necrosis factor (tnf). delineating the sequences in the 14.7k protein that are required for this activity may provide insight into the mechanism of protection from tnf by 14.7k as well as the mechanism of tnf cytolysis. in the present study, we examined the ability of 14.7k mutants to protect cells from lysis by tnf. in-frame deletions as well as c ... | 1993 | 8445725 |
| adenovirus early region 4 and viral dna synthesis. | mutants of human adenovirus type 5 (ad5) lacking early region 4 (e4) display a complex phenotype that includes a delay in the onset of viral dna replication in low-multiplicity infections. studies of viral dna replication in vitro have not revealed a requirement for e4 products in dna synthesis and, for most e4 mutants, defects in dna replication are not apparent at high multiplicities of infection. the effects of e4 mutations on dna replication therefore may reflect a role for e4 in the regulat ... | 1993 | 8460485 |
| a cytoplasmically anchored nuclear protein interferes specifically with the import of nuclear proteins but not u1 snrna. | a cytoplasmically anchored mutant sv40 t antigen, fs t antigen, was shown previously to interfere specifically with the nuclear import of a heterologous nuclear protein, adenovirus 5 fiber protein, in cultured monkey cells (schneider, j., c. schindewolf, k. van zee, and e. fanning. 1988. cell. 54:117-125; van zee, k., f. appel, and e. fanning. 1991. mol. cell. biol. 11:5137-5146). in this report, we demonstrate that fs t antigen also interferes with the nuclear import of adenovirus e1a and a pep ... | 1993 | 8468344 |
| selective effects on adenovirus late gene expression of deleting the e1b 55k protein. | the e1b gene of human adenovirus 5 (ad5) encodes a 55k product previously shown to be required for the efficient accumulation of mrnas derived from the major late primary transcript in the cytoplasm of infected cells. this finding is extended here to include the transcripts from other viral promoters activated during the late phase of infection. conversely, accumulation of mrna derived from the major late promoter at early times is not dependent on this e1b function. cytoplasmic levels of the va ... | 1993 | 8468551 |
| role of phosphorylation near the amino terminus of adenovirus type 5 early region 1a proteins. | human adenovirus early region 1a (e1a) proteins act as transcriptional regulators and function in the control of dna synthesis and cell transformation. little is known about how these viral products are functionally regulated. e1a proteins of adenovirus serotype 5 (ad5) are phosphorylated at several serine residues and previous studies had indicated that both ser-89 and ser-219 are substrates for one or more of the cdc2 family of cell cycle kinases. a second residue near the amino terminus, ser- ... | 1993 | 8468552 |
| high affinity and specificity of dimeric binding of thyroid hormone receptors to dna and their ligand-dependent dissociation. | the thyroid hormone receptors are ligand-dependent, dna binding, trans-acting transcriptional factors belonging to the erba-related steroid/thyroid hormone receptor superfamily. we report here the high affinity and specificity of dimeric dna binding of human thyroid hormone receptor-alpha 1 (htr alpha 1) and htr beta 1 and the effect of t3 on its dna binding. gel mobility shift assay showed that htr alpha 1 and -beta 1 bind to the rat gh-thyroid hormone response element (tre) and rat malic enzym ... | 1993 | 8469235 |
| induction of the cell cycle in baby rat kidney cells by adenovirus type 5 e1a in the absence of e1b and a possible influence of p53. | from previous studies on the induction of dna synthesis in quiescent primary baby rat kidney cells by adenovirus type 5 (ad5) e1a deletion mutants, we concluded that induction is prevented only when cellular proteins p300 and prb are both uncomplexed with e1a (j.a. howe, j.s. mymryk, c. egan, p.e. branton, and s.t. bayley, proc. natl. acad. sci. usa 87:5883-5887, 1990). we have now examined induction by these same mutants in virus lacking the e1b region, so that cellular p53 was no longer comple ... | 1993 | 8474183 |
| a multicomponent cis-activator of transcription of the e1b gene of adenovirus type 5. | we report that transcription of the adenovirus type 5 e1b gene is activated substantially in cis by sequences located between positions -362 and -49 with respect to the rna start site. dna fragments consisting of the -362 to -49 sequences, or subsets thereof, were inserted into a reporter plasmid containing a minimal e1b promoter (positions -48 to +14) joined to the escherichia coli cat gene. in the presence of cotransfected e1a and e1b genes in trans, cat enzyme synthesis in transfected kb cell ... | 1993 | 8480416 |
| expression of the gene encoding secreted placental alkaline phosphatase (seap) by a nondefective adenovirus vector. | a nondefective recombinant human adenovirus 5 (ad5) carrying the seap gene, encoding human secreted placental alkaline phosphatase, in the e3 region of the ad5 genome was obtained. the expression of seap at the early and late stages of ad5 infection was demonstrated in permissive and semi-permissive cell cultures. the amount of seap in the culture medium of the 293 cells was 13.6% of the total protein. | 1993 | 8482541 |
| coevolution of persistently infecting small dna viruses and their hosts linked to host-interactive regulatory domains. | although most rna viral genomes (and related cellular retroposons) can evolve at rates a millionfold greater than that of their host genomes, some of the small dna viruses (polyomaviruses and papillomaviruses) appear to evolve at much slower rates. these dna viruses generally cause host species-specific inapparent primary infections followed by life-long, benign persistent infections. using global progressive sequence alignments for kidney-specific polyomaviridae (mouse, hamster, primate, human) ... | 1993 | 8483926 |
| regulated splicing of adenovirus type 5 e4 transcripts and regulated cytoplasmic accumulation of e4 mrna. | the e4 gene of human type c adenoviruses has been shown previously to give rise to an array of mrnas via differential splicing. in this study, the pattern of expression of these mrnas during lytic infection was examined, and two distinct temporal classes were defined. mrnas of the early class were distinguished from those of the late class by the presence, in the early class, of a sequence in the 3' half of the mrna that was removed as an intron in the late class. a single mrna of the late class ... | 1993 | 8497048 |
| rna-binding properties of a translational activator, the adenovirus l4 100-kilodalton protein. | the adenovirus l4 100-kda nonstructural protein (100k protein) is required for efficient initiation of translation of viral late mrna species during the late mrna species during the late phase of infection (b. w. hayes, g. c. telling, m. m. myat, j. f. williams, and s. j. flint, j. virol. 64:2732-2742, 1990). the rna-binding properties of this protein were analyzed in an immunoprecipitation assay with the 100k-specific monoclonal antibody 2100k-1 (c. l. cepko and p. a. sharp, virology 129:137-15 ... | 1993 | 8497066 |
| p53 mutations in human immortalized epithelial cell lines. | although rodent cells have been immortalized following transfection with a mutant p53 gene, the role of p53 in the immortalization of human cells is unknown. therefore, human epithelial cell lines were examined for p53 mutations in exons 4-9 which include the evolutionarily conserved regions. a spontaneously immortalized skin keratinocyte cell line, hacat, and three ras-transfected clones, have a p53 mutational spectrum that is typical of ultraviolet light induced mutations. a normal finite life ... | 1993 | 8504475 |
| the nfiii/oct-1 binding site stimulates adenovirus dna replication in vivo and is functionally redundant with adjacent sequences. | the inverted terminal repeat (itr) of adenovirus type 5 (ad5) is 103 bp in length and contains the origin of dna replication. cellular transcription factors nfi/ctf and nfiii/oct-1 bind to sites within the itr and participate in the initiation of viral dna replication in vitro. the itr also contains multiple copies of two conserved sequence motifs that bind the cellular transcription factors sp1 and atf. we have analyzed a series of viruses that carry deletions at the left terminus of ad5. a vir ... | 1993 | 8510211 |
| increased cyclin a and decreased cyclin d levels in adenovirus 5 e1a-transformed rodent cell lines. | adenovirus-(ad)- e1a proteins carry two conserved domains (cr1 and cr2) required for transformation of primary rodent cells and essential for association with cellular proteins, including p105rb, p58cyclin a and p33cdk2. we show that in normal rat kidney 49f (nrk) cell lines expressing various mutant ad5-e1a genes, cr2-, but not cr-1-, deletion mutants induce a typical transformed phenotype as characterized by morphology, absence of density arrest and loss of serum requirement. this indicates th ... | 1993 | 8510923 |
| the human papillomavirus type 16 e7 protein complements adenovirus type 5 e1a amino-terminus-dependent transactivation of adenovirus type 5 early genes and increases atf and oct-1 dna binding activity. | we have previously shown that conserved region 1 (cr1) of the adenovirus type 5 (ad5) e1a protein synergizes with cr3 in the transactivation of ad5 early genes (h.k. wong and e. b. ziff, j. virol. 68:4910-4920, 1994). cr1 lies within the e1a amino terminus and binds host regulatory proteins such as the rb protein, p107, p130, and p300. since simian virus 40 (sv40) large t antigen and human papillomavirus type 16 (hpv16) e7 protein also bind host regulatory factors, we investigated whether these ... | 1996 | 8523545 |
| efficient dual transcomplementation of adenovirus e1 and e4 regions from a 293-derived cell line expressing a minimal e4 functional unit. | transgene expression after the administration of recombinant adenovirus with e1 deleted is constantly transient. it is admitted that e1a-substituting activities of cellular or viral origin allow viral antigen synthesis and trigger cytotoxic lymphocyte-mediated clearance of the recipient cells. our approach to solving this problem relies on the additional deletion of the e4 region from the vector backbone as this region upregulates viral gene expression at both transcriptional and posttranscripti ... | 1996 | 8523570 |