| adeno-associated virus in adenovirus type 3 conjunctivitis. | although human infection with adenovirus-associated virus (aav) has been demonstrated, there is no evidence that disease results from such infections. the proportion of adenovirus infections which are dual infections with aav is virtually unknown, since special methods are required to demonstrate infection with aav. to search for aav, we re-examined a collection of specimens from 40 persons involved in an epidemic of pharyngoconjunctival fever associated with a swimming pool. virological and ser ... | 1975 | 166918 |
| effect of adeno-associated virus on cancer expression by herpesvirus-transformed hamster cells. | infection of herpes simplex virus type-2-transformed hamster tumor cells with adeno-associated virus type 1 before inoculation into hamsters specifically delayed the appearance of palpable tumors and increased the survival time of the animals. the data indicated that a defective virus of humans can influence cancer expression by a virus-transformed cell. | 1975 | 171433 |
| structure and nucleotide sequence of the terminal regions of adeno-associated virus dna. | | 1977 | 194395 |
| serological response of chickens exposed to a type 1 avian adenovirus alone or in combination with the adeno-associated virus. | the avian adenoviruses (av) are common infectious agents of poultry and other avian species throughout the world (1,4,8). limited observations suggest that the adeno-associated virus (a-av) coinfects many of the chickens that carry av (8). the presence and persistence of these infections in a flock is often determined by serological methods. in the current study, the immune response of chickens to type 1 av alone and to a dual exposure, av plus a-av, was followed over a 12-week period with a var ... | 1977 | 199153 |
| effect of avian adeno-associated virus on pathogenicity of tipton virus in chicks. | the avian adeno-associated virus (a-av) reduced the pathogenicity of an adenovirus infection in vivo. groups of chicks were infected with tipton virus alone or in combination with high or low doses of a-av. in both trials, the associated virus delayed and reduced chick mortality. this effect was dose-dependent and significant at the higher dose level. | 1978 | 209781 |
| complementation of adeno-associated virus growth with temperature-sensitive mutants of human adenovirus types 12 and 5. | temperature-sensitive mutants of human adenovirus types 12 and 5, defective in viral dna synthesis, were able to support growth of adeno-associated virus type i at the non-permissive temperature. | 1975 | 213533 |
| [helper viruses of adeno-associated virus type 4 replication]. | in replication of adeno-associated virus type 4 (aav-4) the helper function may be performed by a non-defective virus from the same group of parvoviruses (kilham virus). the synthesis of aav-4 antigen was observed in a pig embryo kidney cell line, spev, chronically infected with kilham virus, strain rv-13, 45--52 passages. a one-day-old spev-kilham culture was infected with aav-4. the aav-4 antigen was detected by immunofluorescence at 6, 8, 12, 18 hours, 2, 3, 4, and 5 days after inoculation. d ... | 1979 | 217178 |
| parvovirus rna transcripts containing sequences not present in mature mrna: a method for isolation of putative mrna precursor sequences. | we report here a method of rna preparation that may enrich for precursor rna sequences and the results of an investigation of adeno-associated virus (aav) rna transcription that used this method. whole cells were lysed with detergent and high salt and separated into supernatant and pellet (crude chromatin) fractions. these fractions were then separately deproteinized by proteolytic digestion and phenol extractions. dna was removed from the preparation by two cycles of pancreatic dnase digestion ... | 1979 | 218213 |
| adeno-associated virus autointerference. | | 1979 | 218354 |
| adeno-associated virus dna replication: nonunit-length molecules. | | 1979 | 219605 |
| defective-interfering particles of the human parvovirus adeno-associated virus. | | 1979 | 220782 |
| adeno-associated virus dna replication complexes in herpes simplex virus or adenovirus-infected cells. | a complex which is active in in vitro synthesis of adeno-associated virus (aav) dna was solubilized from vero cells that were co-infected with aav and either adenovirus (ad5) or a herpes simplex virus type 1 (hsv-1) as the helper virus. the complexes from the ad5 and hsv-1-infected cells sedimented at 23 s and 28 s, respectively. the optimal conditions for in vitro dna synthesis for the two types of complex using the endogenous aav template and the endogenous dna polymerase, differed with respec ... | 1979 | 221504 |
| studies on the defectiveness of adeno-associated virus (aav). i. effects of phosphonoacetic acid and 2-deoxy-d-glucose on the replication of aav. | | 1979 | 222062 |
| studies on the defectiveness of adeno-associated virus (aav). ii. effect of growth in cv-1 cells on the replication of adeno-associated virus. | | 1979 | 222063 |
| isolation of an adeno associated virus from sheep. brief report. | an adenovirus and a spherical virus 20--24 nm diameter were isolated from ovine faeces. the small virus replicated in the nucleus, and was associated especially with the nucleolus. it haemagglutinated guinea pig and human erythrocytes, was thermostable and required an adenovirus for replication. it is concluded that this represents the first recorded isolate of an ovine aav. | 1979 | 226038 |
| adeno-associated virus dna replication. | | 1979 | 226321 |
| adeno-associated virus--an extreme state of viral defectiveness. | | 1979 | 228349 |
| helper factor(s) for growth of adeno-associated virus in cells transformed by adenovirus 12. | evidence is presented that a helper factor(s) for growth of adeno-associated virus (aav) is present in cells transformed by adenovirus type 12 (ad12). the growth of aav was observed in heterokaryons formed by fusion of human kb and ad12-transformed rodent cells by using ultraviolet-inactivated sendai virus without coinfection of cells with adenovirus. the presence of the helper factor(s) for aav growth in rat cells transformed by the ecori-c fragment or the hindiii-g fragment of ad12dna suggests ... | 1977 | 337298 |
| detection and serological identification of adeno-associated virus in avian adenovirus stocks. | eleven avian adenovirus strains were tested for the presence of avian adeno-associated viruses (aaav). six strains contained aaav. electron microscopy using rabbit anti-aaav serum was useful in detecting the satellite virus. the aaav previously isolated from guail bronchitis virus was related to each of the six new isolates by immunoagglutination, complement fixation, immunodiffusion, and neutralization tests. | 1975 | 803468 |
| viral dna synthesis in vitro with nuclei isolated from adeno-associated virus type 1-infected cells. | | 1977 | 841869 |
| origin and termination of adeno-associated virus dna replication. | | 1977 | 867815 |
| establishment and characterization of kb cell lines latently infected with adeno-associated virus type 1. | | 1977 | 898681 |
| isolation of the viral dna replication complex from adeno-associated virus type 1-infected cells. | the replication complex active in adeno-associated virus type 1 (aav-1) dna synthesis in vitro was solubilized, with a nonionic detergent, from the nuclei of human embryonic kidney cells coinfected with aav-1 and an early temperative-sensitive mutant (ts125) of human adenovirus type 5 at the nonpermissive temperature (40.5 degrees c). the complex sedimented with a mean size of 23s and contained parental aav-1 dna. most of the dna synthesized with the aav-1 dna replication complex in vitro was aa ... | 1977 | 916024 |
| evidence for two nucleotide sequence orientations within the terminal repetition of adeno-associated virus dna. | duplex adeno-associated virus (aav) dna, produced by annealing plus and minus virion single strands, has been digested with several bacterial restriction endonucleases. these studies reveal the existence of alternate secondary structures at the termini of duplex aav dna. analysis of the sites of endo r-hpa ii cleavage, the products of complete endo r-hpa ii digestion, and the multiple terminal secondary structures leads to the conclusion that there are two possible nucleotide sequences at each e ... | 1977 | 916029 |
| adeno-associated virus dna structure. restriction endonuclease maps and arrangement of terminal sequences. | | 1977 | 919346 |
| intracellular distribution and polyadenylate content of adeno-associated virus rna sequences. | | 1976 | 960561 |
| genome localization of adeno-associated virus rna. | in previous work, linear duplex molecules of adeno-associated virus, type 2 (aav2), dna were cleaved with the restriction endonucleases r-ecori, r-hindii, and r-hindiii. the physical order of the specific fragments obtained was deduced and oriented with respect to the dna strand polarity and the direction of transcription. stable aav rna is transcribed only from 70% of the minus dna strand. we report here rna-dna hybridization experiments using these restriction fragments to obtain a more accura ... | 1976 | 972429 |
| multiplication of adeno-associated virus type 1 in cells coinfected with a temperature-sensitive mutant of human adenovirus type 31. | | 1976 | 982809 |
| isolation and characterization of adeno-associated viruses from bovine adenovirus types 1 and 2. | hemagglutinating dna viruses of 20 nm diameter were isolated from bovine adenovirus types 1 and 2. the isolates were heat stable, chloroform resistant, and defective. their densities were 1.38 to 1.39 g/cm3, and they were found to be serologically identical to the bovine adeno-associated virus strain x7. a partial antigenic relationship was found between these and the canine adeno-associated virus. | 1976 | 986122 |
| cleavage of adeno-associated virus dna with sali,psti and haeii restriction endonucleases. | duplex aav-2 dna was digested with sali, psti or haeii restriction endonucleases and the cleavage sites were mapped. sali cleaves aav dna at 0.310 map units, psti at 0.106, 0.422 and 0.914 and the five haeii sites were mapped at 0.110. 0.156, 0.181, 0.536 and 0.600 map units. these cleavage products will be useful for the isolation of specific regions from the aav dna, located outside of the stably transcribed region of the genome, and will also help to map more complex restriction enzyme cleava ... | 1976 | 995645 |
| study of the fine structure of adeno-associated virus dna with bacterial restriction endonucleases. | a physical map of the adeno-associated virus type 2 genome has been constructed on the basis of the five fragments produced by the restriction endonucleases hindii + iii from hemophilus influenzae. there are three endo r-hindii cleavage sites and one endo r-hindiii site. evidence has been obtained to support the existence of two nucleotide sequence permutations in adeno-associated virus dna, the start points of which have been estimated to be separated by 1% of the genome. the three cleavage fra ... | 1975 | 1159899 |
| [study of the possibilities of inactivating adeno-associated virus type 4]. | adeno-associated type 4 virus (aav-4) is inactivated by formalin dilutions 1 : 400, 1 : 1000 and 1 : 2000 for 24 hours at 37 degrees c. under these conditions hemogglutinins (ha) and complement-fixing antigen (cfa) are retained. the residual formalin must be neutralized with sodium bisulphite. aav-4 can also be inactivated by 3% hydrogen peroxide and ha and cfa are retained too. hydrogen peroxide may be neutralized with catalase. no inactivation of aav-4 could be achieved with lowere concentrati ... | 1975 | 1162954 |
| detection of adeno-associated virus (aav)-specific nucleotide sequences in dna isolated from latently infected detroit 6 cells. | | 1975 | 1198930 |
| [development of a method for preparing adeno-associated virus type 4 antigen]. | a method for preparation of adeno-associated type 4 virus (aav-4) purified from group-specific adenovirus antigen by adsorption on formalinized sheep erythrocytes and elution into hypertonic nacl solution was developed. in 1 m nacl solution the purified aav-4 retained its infectivity and the complement-fixing and hemagglutinating activities. separation of aav-4 and adenovirus group-specific complement-fixing antigen was based on differences in conditions of their adsorption and elution. aav-4 wa ... | 1976 | 1258451 |
| the presence of avian adenoviruses and adeno-associated viruses in healthy chickens. | avian adenoviruses were isolated from 56 of 106 fecal and rectal tissue samples taken from apparently healthy young chickens on 4 farms. only one isolation was made from the 37 samples from 3- and 4-week-old chicks, while the isolation frequency was 64-100% in groups 5 weeks old and older. the 56 adenovirus isolates were classified into 6 serotypes. vurses of 3 or 4 types were found on each farm. avian adeno-associated virus cf antigens were found, using chicken embryos coinfected with an adenov ... | 1976 | 1259656 |
| multiple structures of adeno-associated virus dna: analysis of terminally labeled molecules with endonuclease r-hae iii. | the double-stranded form of adeno-associated virus (aav) dna has about 20 sites sensitive to endonuclease r.hae iii from haemophilus aegypitus; the fragments produced fall into about 13 size classes, 8 of which contain single fragments. the location of the hae iii-produced aav fragments relative to the three ecor1 fragments was determined. using revised figures for the molecular weights of the hae iii cleavage products of phix174 replicative form dna, we calculated that aav dna contains about 4, ... | 1976 | 1271522 |
| cloning of a negative transcription factor that binds to the upstream conserved region of moloney murine leukemia virus. | the long terminal repeat of moloney murine leukemia virus (mulv) contains the upstream conserved region (ucr). the ucr core sequence, cgccatttt, binds a ubiquitous nuclear factor and mediates negative regulation of mulv promoter activity. we have isolated murine cdna clones encoding a protein, referred to as ucrbp, that binds specifically to the ucr core sequence. gel mobility shift assays demonstrate that the ucrbp fusion protein expressed in bacteria binds the ucr core with specificity identic ... | 1992 | 1309593 |
| partial purification of adeno-associated virus rep78, rep52, and rep40 and their biochemical characterization. | we have used differential cell extraction and conventional chromatography to separate and partially purify the four adeno-associated virus (aav) nonstructural proteins rep78, rep68, rep52, and rep40. in the cytoplasmic extracts rep52 and rep40 were present in greater abundance than rep68 and rep78, with rep78 being the least abundant. in nuclear extracts the four rep proteins were approximately equal in abundance. regardless of the subcellular fraction examined, three of the rep proteins (rep78, ... | 1992 | 1309894 |
| in vitro resolution of adeno-associated virus dna hairpin termini by wild-type rep protein is inhibited by a dominant-negative mutant of rep. | an adeno-associated virus (aav) genome with a lys-to-his (k340h) mutation in the consensus nucleotide triphosphate binding site of the rep gene has a dominant-negative dna replication phenotype in vivo. we expressed both wild-type (rep78) and mutant (rep78ntp) proteins in two helper-free expression systems consisting of either recombinant baculoviruses in insect cells or the human immunodeficiency virus type 1 long terminal repeat promoter in human 293 cell transient transfections. we analyzed n ... | 1992 | 1309900 |
| influence of adeno-associated virus on adherence and growth properties of normal cells. | it has been shown previously that infection of newly established cell cultures from malignant human tumors with adeno-associated parvovirus type 2 or type 5 results in growth arrest and cell death. here we report that the additionally observed antiproliferative effect on diploid human fibroblasts is transient and is connected to a reduced number of cells in s phase. progression through the cell cycle is disturbed either in g0/g1 or at the g1/s boundary, but an additional arrest in g2 cannot be e ... | 1992 | 1309911 |
| modification of some biological properties of hela cells containing adeno-associated virus dna integrated into chromosome 17. | parvoviruses are known to interfere with cellular transformation and carcinogenesis. since infecting adeno-associated virus (aav) frequently integrates its dna into the cellular genome, we analyzed whether this integration influences the transformed phenotype of the human tumor cell line hela. analysis of three independent hela cell clones with integrated aav dna (ha-3x, ha-16, and ha-28) revealed the following phenotypic changes of these cells: (i) reduced growth rate, (ii) increased serum requ ... | 1992 | 1313913 |
| rescue of the adeno-associated virus 2 genome correlates with alterations in dna-modifying enzymes in human cells. | the adeno-associated virus 2 (aav) genome can be rescued either from a recombinant plasmid upon transfection into human cells or from cells latently infected with aav, following subsequent infection with adenovirus. using human diploid fibroblasts as a model for a natural aav infection, we observed increased efficiency of rescue of the aav genome in these cells as they traversed their limited proliferative life span in vitro. the efficiency of rescue correlated well with the augmented nuclease a ... | 1992 | 1314783 |
| use of adeno-associated virus as a general transduction vector for mammalian cells. | | 1992 | 1316261 |
| in vitro replication of adeno-associated virus dna. | an in vitro assay for adeno-associated virus (aav) dna replication has been developed. the substrate is a plasmid containing the duplex form of aav dna in pbr322. the aav insert is excised or rescued from the plasmid by extracts of uninfected cells. replication was assayed by production of full-length excised aav dna resistant to dpn i digestion. the following results were obtained. (i) only extracts of cells coinfected with aav and adenovirus replicated the excised insert. (ii) density label ex ... | 1992 | 1316616 |
| gene transfer in human lymphocytes using a vector based on adeno-associated virus. | adeno-associated virus is a nonpathogenic, dependent parvovirus that integrates at a specific site in human chromosome 19. we have used the inverted terminal repeats of the virus, which mediate integration, to establish a vector for gene transfer in human lymphocytes. a neomycin resistance gene has been stably introduced into nontransformed human t-cell clones and a subsequent analysis of the functional properties of the infected clone revealed no detectable alterations. rescue and replication o ... | 1992 | 1318074 |
| analysis of mutations in adeno-associated virus rep protein in vivo and in vitro. | the adeno-associated virus (aav) rep protein is required for both viral dna replication and transactivation of the aav promoters. here we report the effects of mutations in the rep gene on transcription and replication in vivo and terminal repeat binding and terminal resolution site (trs) endonuclease activities in vitro. in all, we examined 10 in-frame deletions and 14 amino acid substitution mutations at eight positions. the point mutations were targeted to regions that are highly conserved am ... | 1992 | 1318396 |
| induction of differentiation-associated changes in established human cells by infection with adeno-associated virus type 2. | the nonpathogenic human defective parvovirus adeno-associated virus (aav) type 2 induced differentiation-associated antigens in cells of the human leukemia cell line hl60 (cd 67), as well as in two different lines of immortalized human keratinocytes, hacat and hpk ia cells (involucrin and cytokeratin 10). simultaneously, expression of the c-myc and c-myb oncogenes and the retinoblastoma gene was down regulated whereas c-fos expression increased in infected cells. these data point to the potentia ... | 1992 | 1318400 |
| inhibition of bovine papillomavirus plasmid dna replication by adeno-associated virus. | the helper-dependent human parvovirus adeno-associated virus type 2 (aav) inhibits both the oncogenic transforming abilities and the dna replication of its helper viruses, adenovirus (ad), and herpes simplex virus (hsv). as aav-2 also inhibits the transforming ability of bovine papillomavirus type 1 (bpv), aav-2 was assayed for its ability to inhibit bpv plasmid dna replication. here we find that the aav-2 rep78 gene is able to trans-inhibit bpv plasmid dna replication and that the aav-2 termina ... | 1992 | 1318608 |
| detection of adeno-associated virus type 2 in human peripheral blood cells. | the non-pathogenic human parvovirus, adeno-associated virus (aav) is helper virus-dependent. however, it integrates into the cellular genome in the absence of its helper viruses. therefore it could become a useful vector for gene therapy. previous studies and our own results have shown that 40 to 80% of adults are seropositive for aav and that seroconversion occurs during the first few years of life, but little is known about the route of natural infection with the virus. we used the polymerase ... | 1992 | 1321880 |
| regulated high level expression of a human gamma-globin gene introduced into erythroid cells by an adeno-associated virus vector. | gene therapy of severe hemoglobinopathies will require high-level expression of a transferred globin gene in erythroid cells. distant regulatory elements flanking the beta-globin gene cluster, the locus control region, are needed for appropriate expression. we have explored the use of a human parvovirus, the adeno-associated virus (aav), for globin gene transfer. the human a gamma-globin gene, linked to hypersensitivity site 2 from the locus control region of the beta-globin gene cluster, was su ... | 1992 | 1323131 |
| adeno-associated virus sensitizes hela cell tumors to gamma rays. | infection with the helper virus-dependent human parvovirus adeno-associated virus (aav) is known to interfere with cellular transformation in vitro and oncogenesis in vivo. here we report on sensitization to gamma irradiation by aav infection of cells in culture and of tumors established from hela cells grafted into immunodeficient (nude) mice: infection of hela cells with aav type 2 enhanced cell killing and reduced plating efficiency after irradiation compared with uninfected cells. similarly, ... | 1992 | 1323717 |
| gene expression from adeno-associated virus vectors in airway epithelial cells. | lung diseases such as cystic fibrosis (cf) might be treated by gene therapy using viral vectors delivered to the airway. one potential vector is the defective human parvovirus, adeno-associated virus (aav). we examined the aav p5 transcription promoter for gene expression in immortalized cell lines derived from the airway (ib3-1) or pancreas (cfpac-1) of cf patients. aav vectors expressing the prokaryotic genes cat (paavp5cat) or neo (paavp5neo) from the p5 promoter were evaluated after introduc ... | 1992 | 1325813 |
| mutational analysis of the adeno-associated virus rep gene. | the replication (rep) gene of the human parvovirus adeno-associated virus (aav) is a pleiotropic effector of numerous viral functions and experts profound effects on cellular transformation. of the four rep proteins, the primarily nuclear rep78 and rep68 direct aav dna replication, trans activation of the capsid (cap) gene promoter, and inhibition of cellular proliferation mediated by various oncogenes. in an initial attempt to define functional domains in rep78, we have constructed a comprehens ... | 1992 | 1326656 |
| modulation of the cellular phenotype by integrated adeno-associated virus. | the adeno-associated virus (aav) rep gene encodes a series of overlapping, multifunctional, nonstructural proteins (rep proteins) which regulate the viral life cycle and which are also capable of trans-regulating nonviral gene expressions (reviewed in berns, 1990, microbiol. rev. 54, 316-329). to investigate the expression of the aav rep gene in a cellular chromosomal context, sv40-transformed chinese hamster embryo (od4) cells were infected with an aav/neo hybrid virus and progeny resistant to ... | 1992 | 1326806 |
| cloning and integration of dna fragments in human cells via the inverted terminal repeats of the adeno-associated virus 2 genome. | in current systems for molecular cloning of eukaryotic genes, bacterial cells are routinely utilized as intermediate hosts. we investigated the possibility of using a viral system for cloning dna fragments independent of bacterial cell usage. in this report, we provide an alternative approach for molecular cloning of dna fragments in eukaryotic cells by utilizing the inverted terminal repeats (itrs) of the genome of a nonpathogenic human parvovirus, the adeno-associated virus 2 (aav). we constru ... | 1992 | 1327964 |
| adenovirus containing a deletion of the early region 2a gene allows growth of adeno-associated virus with decreased efficiency. | efficient growth of adeno-associated virus (aav) requires helper functions provided by a coinfecting adenovirus or herpesvirus. earlier studies using adenoviruses having temperature-sensitive lesions in the early region 2a gene (e2a) produced contradictory evidence regarding the role of the e2a 72-kda dna-binding protein (dbp) in allowing efficient aav growth. these disparate results may reflect varying levels of residual function in the temperature-sensitive dbp. we examined this issue using an ... | 1992 | 1329332 |
| identification of a negative regulatory domain in the human papillomavirus type 18 promoter: interaction with the transcriptional repressor yy1. | the human papillomavirus type 18 (hpv-18) promoter contains a tpa responsive element (tre) which confers tpa responsiveness on a heterologous promoter. in the context of the hpv-18 promoter, however, this ap-1 site is inactive. we have identified a negative regulatory domain in the hpv-18 promoter which represses the constitutive and tpa-induced ap-1 activity. this negative regulatory sequence has been mapped to 44 nucleotides (ol13). we identified this element as a transcriptional silencer base ... | 1992 | 1330541 |
| adeno-associated virus type 2-mediated inhibition of human immunodeficiency virus type 1 (hiv-1) replication: involvement of p78rep/p68rep and the hiv-1 long terminal repeat. | microinjection of wild-type adeno-associated virus type 2 (aav-2) dna and infectious human immunodeficiency virus type 1 (hiv-1) proviral dna into the nuclei of human epithelioid sw480 cells leads to specific inhibition of hiv-1 replication. mutational analysis of the aav genome showed that this negative interference can be assigned to a functional aav-2 rep gene. moreover, the p78rep/p68rep proteins are sufficient for the anti-hiv-1 effects. the rep gene also inhibits the expression of a chlora ... | 1992 | 1331299 |
| assembly of viruslike particles by recombinant structural proteins of adeno-associated virus type 2 in insect cells. | the three capsid proteins vp1, vp2, and vp3 of the adeno-associated virus type 2 (aav-2) are encoded by overlapping sequences of the same open reading frame. separate expression of these proteins by recombinant baculoviruses in insect cells was achieved by mutation of the internal translation initiation codons. coexpression of vp1 and vp2, vp2 and vp3, and all three capsid proteins and the expression of vp2 alone in sf9 cells resulted in the production of viruslike particles resembling empty cap ... | 1992 | 1331503 |
| characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus dna by non-homologous recombination. | the human parvovirus, adeno-associated virus (aav), has been shown to integrate preferentially into human chromosome 19 q13.3-qter. the human target sequence for aav integration (aavs1) was cloned and sequenced. by analysis of the proviral junctions it was determined that integration of the aav dna occurred via a non-homologous recombination pathway although there were either four or five identical nucleotides at the junctions. integration was a multistep, concerted process that resulted in cell ... | 1992 | 1334463 |
| dual-target inhibition of hiv-1 in vitro by means of an adeno-associated virus antisense vector. | an adeno-associated virus vector encoding an antisense rna was used to transduce stable intracellular resistance to human immunodeficiency virus-1 (hiv-1) in human hemopoietic and non-hemopoietic cell lines. the antisense targets are present in all hiv-1 transcripts and include the tar sequence, which is critical for transcription and virus replication, and the polyadenylation signal. cell lines expressing antisense rna showed up to 95 percent inhibition of gene expression directed by the hiv-1 ... | 1992 | 1359646 |
| adeno-associated virus vectors. | adeno-associated virus is a human parvovirus that integrates its dna genome into host cell chromosomes with very high efficiency. this suggests that adeno-associated virus may be a useful vector for human gene therapy. interest in adeno-associated virus vectors increased greatly in the last year following reports that adeno-associated virus genome integration may be site specific and occur at preferred sites in the human genome. several genes relevant to the treatment of genetic or infectious di ... | 1992 | 1369403 |
| colocalization of adeno-associated virus rep and capsid proteins in the nuclei of infected cells. | the mechanism of adeno-associated virus (aav) dna replication was characterized both genetically and biochemically. in this study, we used monoclonal and polyclonal antibodies to examine the aav p5 (rep78 and rep68) and p19 (rep52 and rep40) proteins in infected cells. by overexpressing a truncated rep78 protein in escherichia coli, we obtained monoclonal antibody anti-78/68, which is specific for the p5 rep proteins, and monoclonal antibody anti-52/40, which recognized both the p5 and p19 rep p ... | 1992 | 1370086 |
| replication of adeno-associated virus type 2 in human lymphocytic cells and interaction with hiv-1. | adeno-associated virus (aav) is a nonpathogenic parvovirus which normally requires helper adenovirus or herpes-virus for replication. we examined the growth of aav type 2 in human lymphocytes and its possible interaction with hiv-1. three b cell lines (ck-b, hs-2, and uc729) and four t cell lines (molt-4, jurkat, hut78, and hut78+hiv, which is persistently infected with hiv-1) were infected with aav either in the presence or in the absence of adenovirus. aav dna was found in cells of all the lin ... | 1992 | 1372138 |
| defective regulation of outwardly rectifying cl- channels by protein kinase a corrected by insertion of cftr. | cystic fibrosis (cf) is a lethal genetic disease resulting in a reduced cl- permeability, increased mucous sulphation, increased na+ absorption and defective acidification of lysosomal vesicles. the cf gene encodes a protein (the cystic fibrosis transmembrane conductance regulator, cftr) that can function as a low-conductance cl- channel with a linear current-voltage relationship whose regulation is defective in cf patients. larger conductance, outwardly rectifying cl- channels are also defectiv ... | 1992 | 1380129 |
| chromosomal localization and organization of the murine genes encoding the beta subunits (aic2a and aic2b) of the interleukin 3, granulocyte/macrophage colony-stimulating factor, and interleukin 5 receptors. | chromosomal genes for two mouse homologous beta subunits (aic2a and aic2b) of the interleukin-3, granulocyte/macrophage colony-stimulating factor, and interleukin-5 receptors were characterized. both aic2a and aic2b genes were present on a 250-kilobase mlui restriction fragment and were mapped on murine chromosome 15 (these loci were provisionally designated as il3rb-1 (aic2a) and il3rb-2 (aic2b)), closely linked to the c-sis locus. both genes consist of 14 exons and span about 28 kb each. the m ... | 1992 | 1386365 |
| definition of interferon gamma-response elements in a novel human fc gamma receptor gene (fc gamma rib) and characterization of the gene structure. | the human fc gamma ri (cd64) is a high affinity receptor for the fc portion of immunoglobulin (ig), and its constitutively low expression on the cell surface of monocyte/macrophage and neutrophils is selectively upregulated by interferon gamma (ifn-gamma) treatment (perussia, b., e. t. dayton, r. lazarus, v. fanning, and g. trinchieri. 1983. j. exp. med. 158:1092). three distinct cdnas have been cloned and code for proteins that predict three extracellular ig-like domains (allen, j.m., and b. se ... | 1992 | 1402657 |
| parvoviruses are inefficient in inducing interferon-beta, tumor necrosis factor-alpha, or interleukin-6 in mammalian cells. | to investigate a possible role of cytokines in parvovirus-mediated suppression of tumorigenesis, we tested in cell culture whether parvoviruses are able to induce interferon (ifn)-beta, tumor necrosis factor (tnf)-alpha or interleukin-6 (il-6). infection of rodent or human cells with the parvoviruses minute virus of mice (mvm), h-1 or adeno-associated virus (aav) types 2 or 5 failed to induce expression of the luciferase or beta-galactosidase reporter genes transfected into these cells as constr ... | 1992 | 1522825 |
| inhibition of h-ras expression by the adeno-associated virus rep78 transformation suppressor gene product. | here the adeno-associated virus rep78 gene product was found to inhibit the expression of the chloramphenicol acetyltransferase and the bladder cancer-derived ej-h-ras coding sequences when they were under the control of the natural cellular h-ras regulatory sequences. however, rep78 had little or no effect on the expression of these same coding sequences when they were under the control of the regulatory sequences of the murine osteosarcoma virus long terminal repeat. these data indicate that t ... | 1991 | 1647267 |
| the evolution of defective and autonomous parvoviruses. | because of the small size and genetic simplicity of small dna viruses, parvoviruses would appear to be excellent models for studying viral evolution and adaptation. in an earlier publication we hypothesized the evolution of sequences of cellular "junk" dna into protective interfering transposons. these transposons would interfere with invading pathogenic viruses by competing with the pathogen dna for replicative enzymes. we speculated that a small, defective parvovirus, the adeno-associated viru ... | 1991 | 1648152 |
| adeno-associated virus rep proteins produced in insect and mammalian expression systems: wild-type and dominant-negative mutant proteins bind to the viral replication origin. | the adeno-associated virus (aav) rep gene proteins, rep78 and rep68, are required for replication of aav dna and bind to the aav replication origin. an aav genome having a lys340 to his (k340h) mutation in the consensus purine nucleotide binding site of the rep gene protein exhibited a dominant-negative phenotype for dna replication. we synthesized both wild-type and the k340h mutant rep78 protein in a baculovirus expression system. nuclear extracts of sf9 cells containing these proteins were ex ... | 1991 | 1651588 |
| adeno-associated viruses having nonsense mutations in the capsid genes: growth in mammalian cells containing an inducible amber suppressor. | when an adeno-associated virus (aav) genome contained in a recombinant plasmid is transfected into adenovirus-infected cells, infectious aav particles are efficiently generated. we previously described the construction of a conditional lethal mutant of aav having an amber termination codon inserted in the rep gene. this mutant was propagated on a monkey kidney cell line (supd12) having an inducible amber suppressor trnaser. we now describe the construction and propagation of two additional condi ... | 1991 | 1651593 |
| mapping and direct visualization of a region-specific viral dna integration site on chromosome 19q13-qter. | a human parvovirus, adeno-associated virus (aav), is unique among eukaryotic dna viruses in its ability to integrate site specifically into a defined region of human chromosome 19. in this study we used in situ hybridization to visualized directly the site of aav dna integration in latently infected human cell lines and normal human cells. | 1991 | 1653762 |
| transcriptional repression by yy1, a human gli-krüppel-related protein, and relief of repression by adenovirus e1a protein. | a sequence within the transcription control region of the adeno-associated virus p5 promoter has been shown to mediate transcriptional activation by the adenovirus e1a protein. we report here that this same element mediates transcriptional repression in the absence of e1a. two cellular proteins have been found to bind to overlapping regions within this sequence element. one of these proteins, yy1, is responsible for the repression. e1a relieves repression exerted by yy1 and further activates tra ... | 1991 | 1655281 |
| construction and expression of a recombinant adeno-associated virus that harbors a human beta-globin-encoding cdna. | towards a goal of using recombinant adeno-associated viruses (aav) for the gene therapy of hemoglobinopathies we had previously constructed plasmid pav h beta g psi 1, which contained a human beta-globin-encoding cdna (hbb) downstream from the p40 promoter of aav2 dna [ohi et al., gene 89 (1990) 279-282]. transfection of the plasmid into human 293 cells (embryonal kidney cell line) resulted in the expression of hbb at the mrna level as well as rescue and replication of the recombinant aav genome ... | 1991 | 1655580 |
| biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus. | two infectious components with buoyant densities of 1.40 g/cm3 and 1.45 g/cm3, designated as major (1.40) and minor (1.45) component, were detected by banding avian adeno-associated virus (aaav) isopycnically in cscl. in metrizamide, however, infectious aaav banded only as a single peak at a density of 1.32 g/cm3. biological as well as physicochemical properties of the two aaav components recovered from cscl density gradient were described. concerning the minor (1.45) component, three experiment ... | 1991 | 1656917 |
| targeted integration of adeno-associated virus (aav) into human chromosome 19. | a key feature in adeno-associated virus (aav) replication is efficient integration of the viral genome into host cell dna to establish latency when helper virus is absent. the steps involved in this process remain largely uncharacterized, even though aav integration was first documented 20 years ago. using a protein--dna binding method we isolated aav--cellular junction dna sequences. the cellular component hybridized to a single restriction fragment in the virus-free parental cell line, and als ... | 1991 | 1657596 |
| replication of the human immunodeficiency virus type 1 is inhibited by the adeno-associated virus rep gene. | | 1991 | 1665443 |
| yy1 is an initiator sequence-binding protein that directs and activates transcription in vitro. | regulation of eukaryotic messenger rna transcription is governed by dna sequence elements that serve as binding sites for sequence-specific transcription factors. these include upstream and downstream promoter-proximal elements, enhancers, repressors, and silencers, which modulate the rate of specific initiation by rna polymerase ii. in addition, the promoter-proximal region between -45 and +30 (relative to the start of initiation) contains two highly conserved motifs, the tata sequence at aroun ... | 1991 | 1720509 |
| nucleotide sequence and genomic organization of aedes densonucleosis virus. | over 99% of the genome of aedes densonucleosis virus was determined. two types of the viral dna were found that differ only in four nucleotides (nt) in the 5' noncoding part and whose sizes are 4009 nt (more copious) and 4012 nt, respectively. both 146 nt at the 3' end and 164 nt at the 5' end could assume a similar t-shaped structure; but unlike the adeno-associated virus, aedes dna has a unique primary dna sequence at each terminus. however, the crossarms of these structures are built of the s ... | 1991 | 1833875 |
| adeno-associated virus rep protein inhibits human immunodeficiency virus type 1 production in human cells. | the adeno-associated virus (aav) rep gene encodes four proteins (rep78, rep68, rep52, and rep40) required for aav dna replication and aav gene regulation. in addition, the rep proteins may have pleiotropic regulatory effects in heterologous systems, and in particular rep78 may mediate a negative regulatory effect. we analyzed the effects of the aav rep gene on human immunodeficiency virus type 1 (hiv-1) gene expression. the rep gene proteins of aav type 2 (aav2) inhibited the trans-activating ab ... | 1991 | 1845899 |
| inhibition of cellular transformation by the adeno-associated virus rep gene. | the replication, or rep, gene of the human parvovirus, adeno-associated virus (aav), is a pleiotropic effector of numerous viral functions. the rep gene trans-regulates viral dna replication, mrna transcription, and assembly of the infectious virion. in addition to its roles in the virus life cycle the rep gene also represses gene expression from viral or cellular transcription promoters in both transient and long-term assays. in this report we have investigated the ability of the rep gene to in ... | 1991 | 1849683 |
| a subset of herpes simplex virus replication genes provides helper functions for productive adeno-associated virus replication. | herpesviruses are helper viruses for productive adeno-associated virus (aav) replication. to analyze the herpes simplex virus type 1 (hsv-1) functions mediating helper activity, we coinfected hela cells with aav type 2 (aav-2) and different hsv-1 mutants defective in individual hsv replication genes. aav replication was fully accomplished in the absence of hsv dna replication and thus did not require expression of late hsv genes. in addition, hsv mutants lacking either the origin-binding protein ... | 1991 | 1850024 |
| in vitro rescue of an integrated hybrid adeno-associated virus/simian virus 40 genome. | in an in vitro simian virus 40 (sv40) dna replication assay, we have observed excision of a hybrid adeno-associated virus (aav)/sv40 insert from a plasmid construct. the excision was dependent on the presence of the palindromic aav terminal repeat and greatly enhanced by the addition of the sv40 t antigen to the reaction. analysis of the excision product supports a model in which the palindromic terminal sequences of aav form a cruciform structure (equivalent to a holliday recombination intermed ... | 1991 | 1850805 |
| acquisition of the human adeno-associated virus type-2 rep gene by human herpesvirus type-6. | human herpesvirus type-6 (hhv-6) is a recently isolated herpesvirus which is highly prevalent in adult populations around the world. hhv-6 was first isolated from the peripheral blood of six individuals with lymphoproliferative disorders, two of whom were also infected with human immunodeficiency virus. hhv-6, in common with other herpesviruses, transactivates the hiv long terminal repeat linked to reporter genes and has in addition been shown to accelerate hiv gene expression and cd4 cell death ... | 1991 | 1851252 |
| conserved chromatin structure in c-myc 5'flanking dna after viral transduction. | the role of local sequence information in establishing the chromatin structure of the human c-myc upstream region (mur) was investigated. adeno-associated virus (aav)-mediated gene transduction was used to introduce an additional unrearranged copy of the 2.4 kb hindiii-xhoi fragment of the mur into a novel location in the genome in each of two cloned hela cell lines. the aav-based rep- cap- viral vector skma used to transduce the mur retained only 1.4 kb (24%) of the aav genome and could accommo ... | 1991 | 1942068 |
| isolation of a candidate repressor/activator, nf-e1 (yy-1, delta), that binds to the immunoglobulin kappa 3' enhancer and the immunoglobulin heavy-chain mu e1 site. | we have determined that the developmental control of immunoglobulin kappa 3' enhancer (kappa e3') activity is the result of the combined influence of positive- and negative-acting elements. we show that a central core in the kappa e3' enhancer is active at the pre-b-cell stage but is repressed by flanking negative-acting elements. the negative-acting sequences repress enhancer activity in a position- and orientation-independent manner at the pre-b-cell stage. we have isolated a human cdna clone ... | 1991 | 1946405 |
| sequences required for coordinate induction of adeno-associated virus p19 and p40 promoters by rep protein. | a series of contiguous 30-bp deletions were introduced into the regions upstream of the p19 and p40 promoters of adeno-associated virus (aav), and the effects of these deletions on induction of aav transcription by the rep gene products was evaluated. a novel complementation system was devised for supplying wild-type rep protein when mutations disrupted the trans activation activity of the rep protein. transcription from the p40 promoter was eliminated upon deletion of the tata sequence located ... | 1991 | 2033660 |
| origin of adeno-associated virus dna replication is a target of carcinogen-inducible dna amplification. | dna amplification of the helper-dependent parvovirus aav (adeno-associated virus) can be induced by a variety of genotoxic agents in the absence of coinfecting helper virus. here we investigated whether the origin of aav type 2 dna replication cloned into a plasmid is sufficient to promote replication activity in cells treated by the carcinogen n-methyl-n'-nitro-n-nitrosoguanidine (mnng). a puc19-based plasmid, designated pa2y1, which contains the left terminal repeat sequences (trs) representin ... | 1991 | 2033669 |
| parvovirus b19-induced perturbation of human megakaryocytopoiesis in vitro. | parvovirus b19 infection leads to transient aplastic crises in individuals with chronic hemolytic anemias or immunodeficiency states. an additional unexplained sequela of b19 infection is thrombocytopenia. because b19 is known to have a remarkable tropism for human erythropoietic elements, and is not known to replicate in nonerythroid cells, the etiology of this thrombocytopenia is uncertain. we sought to define the pathobiology of b19-associated thrombocytopenia by examining the role of b19 on ... | 1990 | 2146978 |
| in vitro resolution of covalently joined aav chromosome ends. | we have developed an assay for a key step in the replication of adeno-associated virus (aav) dna. we demonstrate the covalently joined ends of linear aav dna can be resolved in vitro to the open duplex configuration. only extracts prepared from human cells that have been infected with both adenovirus and aav are capable of carrying out the reaction. the reaction is initiated by a site-specific and strand-specific endonucleolytic cut at a terminal resolution site near the end of the aav terminal ... | 1990 | 2153052 |
| site-specific integration by adeno-associated virus. | cellular sequences flanking integrated copies of the adeno-associated virus (aav) genome were isolated from a latently infected clonal human cell line and used to probe genomic blots derived from an additional 21 independently derived clones of human cells latently infected with aav. in genomic blots of uninfected human cell lines and of primary human tissue, each flanking-sequence probe hybridized to unique bands, but in 15 of the 22 latently infected clones the flanking sequences hybridized no ... | 1990 | 2156265 |
| mutation of a consensus purine nucleotide binding site in the adeno-associated virus rep gene generates a dominant negative phenotype for dna replication. | adeno-associated virus (aav) contains a multifunctional nonstructural gene, rep, which is required for aav dna replication and has pleiotropic effects on positive and negative regulation of gene expression. all of the parvovirus nonstructural genes contain a region of highly conserved amino acid homology. within this conserved region is the consensus sequence for a purine nucleotide binding site. we constructed a mutant aav having a mutation in this site by converting lysine 340 to histidine. th ... | 1990 | 2157057 |
| the adenovirus dna-binding protein stimulates the rate of transcription directed by adenovirus and adeno-associated virus promoters. | the dna-binding protein (dbp) encoded by the e2a region of adenovirus type 5 was found to enhance the expression of a reporter gene controlled by several different promoters within transfected cells. the rate of synthesis of correctly initiated transcripts was increased by the dbp. the adeno-associated virus p5 promoter and the adenovirus e1a and e2a early and major late promoters responded to the dbp by increases in expression ranging from 6- to 27-fold, while the adenovirus e4 promoter was sli ... | 1990 | 2157873 |
| the aav origin binding protein rep68 is an atp-dependent site-specific endonuclease with dna helicase activity. | genetic studies of adeno-associated virus (aav) indicate that two aav genes are required for viral dna replication: the palindromic terminal repeat, which is the origin for dna replication, and the rep gene, which codes for a family of at least four viral nonstructural proteins. to determine the biochemical function of the rep proteins, we have purified the aav rep68 protein to apparent homogeneity. we find that it contains a site-specific and strand-specific endonuclease activity that specifica ... | 1990 | 2159383 |
| the adeno-associated virus rep gene suppresses herpes simplex virus-induced dna amplification. | herpes simplex virus (hsv) induces within the host cell genome dna amplification which can be suppressed by coinfection with adeno-associated virus (aav). to characterize the aav functions mediating this effect, cloned aav type 2 wild-type or mutant genomes were transfected into simian virus 40 (sv40)-transformed hamster cells together with the six hsv replication genes (encoding ul5, ul8, major dna-binding protein, dna polymerase, ul42, and ul52) which together are necessary and sufficient for ... | 1990 | 2159559 |
| inhibition of n-methyl-n'-nitro-n-nitrosoguanidine-induced methotrexate and adriamycin resistance in cho cells by adeno-associated virus type 2. | we studied the effects of helper-dependent parvovirus aav [adeno-associated virus] type 2 on carcinogen-inducible resistance to methotrexate (mtx) and adriamycin (adr) in chinese hamster ovary cells. both types of drug resistance were monitored by determination of the number of drug-resistant colonies normalized for the respective value of plating efficiency under non-selective conditions. treatment of cells with n-methyl-n'-nitro-n-nitrosoguanidine (mnng) drastically enhanced the frequency of r ... | 1990 | 2161803 |
| construction and replication of an adeno-associated virus expression vector that contains human beta-globin cdna. | with the goal of using adeno-associated viruses (aav) as the gene-transfer vector for gene therapy of hemoglobinopathies, human beta-globin cdna was ligated downstream from the p40 promoter of the aav type-2 (aav2) genome. to circumvent difficulties of cloning dna containing palindromic sequences, two of which exist in the termini of aav genome, a step-wise approach handling one palindrome at a time was devised to construct the chimeric expression vector. electroporation of the construct into hu ... | 1990 | 2165023 |
| infection with adeno-associated virus type 5 inhibits mutagenicity of herpes simplex virus type 1 or 4-nitroquinoline-1-oxide. | infection with adeno-associated virus type 5 (aav-5) reduced the number of mutants arising in the hypoxanthine phosphoribosyltransferase locus of human rd 176 cells after infection with herpes simplex virus type 1 (hsv-1; partially inactivated) or 4-nitroquinoline-1-oxide (4-nqo). the mutation frequency was reduced by aav-5 infection from 11.4 to 1.8 after mutation with hsv-1 and from 3.2 to 2.5 when mutation was induced by 4-nqo. this was analyzed by determination of the number of cells resista ... | 1990 | 2166909 |
| induction of selective dna amplification and morphological cell transformation in syrian hamster embryo cells. | dna amplification is a frequently observed event in continuous cell lines and in tumors. it is likely that a common mechanism underlies the amplification of specific dna sequences which confer drug resistance and genes which give a growth advantage to the tumor. to find a correlation between the induction of dna amplification by chemicals and morphological cell transformation we treated syrian hamster embryo (she) cells with diverse antineoplastic agents of different classes. analysis of these a ... | 1990 | 2170834 |