| 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 |
| 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 |
| physical map and strand polarity of specific fragments of adenovirus-associated virus dna produced by endonuclease r-ecori. | cleavage of adenovirus-associated virus type 2 (aav2) dna linear duplex monomers with the restriction endonuclease r-ecori yielded three fragments, a, b, and c, having approximate mol wt of 1.6 x 10(6), 1.1 x 10(6), and 1.3 x 10(5), respectively. radioactive labeling the 5' termini of aav dna before cleavage with r-ecori showed that a and b were terminal fragments and c was internal. separation of the complementary strands of fragments a and b showed that a contained the 5' terminus of the minus ... | 1975 | 1159894 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| the recombinant human parvoviruses for gene therapy of hemoglobinopathies. | towards a goal of using adeno-associated viruses (aav), the human parvovirus, as the gene transfer vector for gene therapy of hemoglobinopathies, the human beta-globin (h beta g) cdna was ligated downstream of the p40 promoter of aav type 2 (aav2) genome. transfection via electroporation of the construct into human 293 cells (embryonal kidney cell line) resulted in expression of the cloned h beta g cdna, as evidenced by the synthesis of transcripts hybridizable to h beta g probe. the transfectio ... | 1990 | 1366418 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
| adeno-associated virus dna replication is induced by genes that are essential for hsv-1 dna synthesis. | adeno-associated virus (aav) dna replication is not detectable unless cells are coinfected with a helper adenovirus (ad) or herpesvirus or unless aav infection is carried out in certain established cell lines that have been treated with various metabolic inhibitors or uv irradiation. in helper-dependent infections, it has been shown that aav dna synthesis depends on one or more early ad genes, whereas little is known concerning any herpesvirus gene that promotes aav dna synthesis. in this study ... | 1990 | 2173256 |
| efficient synthesis of adeno-associated virus structural proteins requires both adenovirus dna binding protein and va i rna. | we have shown previously that replication of defective parvoviruses [adeno-associated viruses (aav)] requires several early adenovirus (ad) gene products [j. e. janik, m. m. huston, and j. a. rose (1981) proc. natl. acad. sci. usa 78, 1925-1929]. to examine their possible roles in the transcription and translation of aav mrna, 293-31 cells, a human embryonic kidney cell line that constitutively expresses the ad early region ia and ib gene products, were transfected with a pbr325 plasmid (plh1) t ... | 1989 | 2536986 |
| rescue and replication of the adeno-associated virus 2 genome in mortal and immortal human cells. | productive infection of the adeno-associated virus (aav) as well as rescue of the aav genome from a recombinant plasmid and its subsequent replication in established cell lines are dependent upon coinfection with adenovirus. a comparative study of aav infection and rescue/replication was carried out in mortal and immortal human cells. aav dna replication as well as assembly and release of the progeny virions were significantly delayed in normal diploid cells, which have a limited replicative pot ... | 1989 | 2542184 |
| identification of nuclear proteins that specifically interact with adeno-associated virus type 2 inverted terminal repeat hairpin dna. | a palindromic hairpin duplex containing the inverted terminal repeat sequence of adeno-associated virus type 2 (aav) dna was used as a substrate in gel retardation assays to detect putative proteins that specifically interact with the aav hairpin dna structures. nuclear proteins were detected in extracts prepared from human kb cells coinfected with aav and adenovirus type 2 that interacted with the hairpin duplex but not in nuclear extracts prepared from uninfected, aav-infected, or adenovirus t ... | 1989 | 2542611 |
| the adeno-associated virus rep78 gene inhibits cellular transformation induced by bovine papillomavirus. | adeno-associated virus type 2 (aav) is a helper dependent parvovirus which can inhibit the oncogenic and transforming potential of its helper viruses: adenoviruses (ad) and herpesviruses. here we have assayed aav's ability to inhibit cellular transformation induced by bovine papillomavirus type 1 (bpv-1), a member of another family of dna viruses. aav was able to markedly inhibit bpv-1-induced transformation of contact-inhibited murine fibroblasts either by infection with virus particles or by d ... | 1989 | 2549714 |
| construction of a recombinant human parvovirus b19: adeno-associated virus 2 (aav) dna inverted terminal repeats are functional in an aav-b19 hybrid virus. | to facilitate genetic analysis of the human pathogenic parvovirus b19, we constructed a hybrid b19 viral genome in which the defective b19 inverted terminal repeats were replaced with the full-length inverted terminal repeats from a nonpathogenic human parvovirus, the adeno-associated virus 2 (aav). the hybrid aav-b19 genome was rescued from a recombinant plasmid and then the dna was replicated upon transfection into adenovirus 2-infected human kb cells in the presence of aav genes coding for pr ... | 1989 | 2554300 |
| synthesis of adeno-associated virus structural proteins requires both alternative mrna splicing and alternative initiations from a single transcript. | the three adeno-associated virus type 2 (aav2) structural proteins (a, b, and c) are specified by transcripts generated from the most-rightward promoter (p40). protein c (60 kilodaltons [kda]), the most abundantly produced, is entirely contained within b (72 kda) which, in turn, is contained within a (90 kda). although neither of the known structures of p40 transcripts, an unspliced 2.6-kilobase (kb) rna and a spliced 2.3-kb rna, possesses an aug-initiated open reading frame that accounts for th ... | 1988 | 2839699 |
| structure of simian virus 40-adeno-associated virus recombinant genomes. | the structures of recombinant genomes formed by recombination between simian virus 40 (sv40) and adeno-associated virus 2 (aav) dnas after either dna cotransfection or coinfection by virions were characterized. two types of structures were found. group a structures, found after cotransfection and in one of seven recombinants arising from coinfection, represented a simple deletion of sv40 sequences replaced by a slightly shorter aav sequence. group b structures were found in six of seven recombin ... | 1985 | 2997468 |
| direct mapping of adeno-associated virus capsid proteins b and c: a possible acg initiation codon. | the three major capsid proteins of adeno-associated virus type 2 (aav2) virions are designated a, b, and c and have molecular sizes of 90, 72, and 60 kda, respectively. these proteins are related, and genetic studies have shown they are encoded by a long open reading frame located in the right half of the genome. the coding capacity distal to the first atg in this reading frame is only 503 amino acids (i.e., a protein about the size of protein c), but an open frame sequence devoid of atg codons ... | 1985 | 2999784 |
| positive and negative autoregulation of the adeno-associated virus type 2 genome. | the defective human parvovirus, adeno-associated virus (aav), requires multiple functions provided by a coinfecting helper virus for viral replication. in addition, it has recently been shown that at least one aav gene is also required for aav dna replication. in this paper, we investigate the autoregulation of the aav genome by analyzing the expression of mutant aav genomes upon transfection into adenovirus-infected human cells. evidence is presented which indicates that the aav genome regulate ... | 1986 | 3018288 |
| latent infection of kb cells with adeno-associated virus type 2. | adeno-associated virus (aav) is a prevalent human virus whose replication requires factors provided by a coinfecting helper virus. aav can establish latent infections in vitro by integration of the aav genome into cellular dna. to study the process of integration as well as the rescue of aav replication in latently infected cells after superinfection with a helper virus, we established a panel of independently derived latently infected cell clones. kb cells were infected with a high multiplicity ... | 1986 | 3021985 |
| genetics of adeno-associated virus: isolation and preliminary characterization of adeno-associated virus type 2 mutants. | we constructed insertion and deletion mutants with mutations within the adeno-associated virus (aav) sequences of the infectious recombinant plasmid psm620. studies of these mutants revealed at least three aav phenotypes. mutants with mutations between 11 and 42 map units were partially or completely defective for rescue and replication of the aav sequences from the recombinant plasmids (rep mutants). the mutants could be complemented by mutants with replication-positive phenotypes. the protein( ... | 1984 | 6086948 |
| adeno-associated virus proteins: origin of the capsid components. | the three primary capsid proteins (a, b, and c) of adeno-associated viruses have been shown previously to contain overlapping amino acid sequences (r. mcpherson and j. rose, j. virol. 46:523-529, 1983). in the present study we demonstrate definitively that these proteins are encoded in the right half of the adeno-associated virus 2 genome, and one or both of the smallest adeno-associated rna species (2.3- or 2.6-kilobase rna) account for their synthesis. protein a (90 kilodaltons) apparently ini ... | 1984 | 6092680 |
| nucleotide sequence of the inverted terminal repetition in adeno-associated virus dna. | the inverted terminal repetition in adeno-associated virus type 2 dna has been sequenced. the terminal repetition contain 145 nucleotides of which the first 125 nucleotides can self-base pair to form a t-shaped hairpin structure. both restriction endonuclease analysis with smai and bgli and direct sequence analysis of the smai fragments provide evidence for two sequences in the region of the terminal repetition between nucleotides 44 and 81. the two sequences represent an inversion of the first ... | 1980 | 6246271 |
| definition of a novel promoter for the major adenovirus-associated virus mrna. | a 660 nucleotide adenovirus-associated virus type 2 (aav2) dna fragment which encodes the 5' terminal leader and the entire intervening sequence of the major viral mrna has been cloned into pbr322, and its primary sequence has been determined. the 5' terminal viral mrna sequence was deduced by sequencing the reverse-transcriptase cdna extension product of a 5' end-labeled dna primer complementary to the rna 5' terminal region. from combined dna and rna sequence analyses (which confirm our previo ... | 1980 | 6253077 |
| transcripts of the adeno-associated virus genome: mapping of the major rnas. | the four major adeno-associated virus type 2 (aav2)-specific rnas were mapped on the linear viral genome by a variety of biochemical techniques, including s1 nuclease and exonuclease vii mapping, rna gel-transfer hybridization, and analysis of reverse transcriptase extension products. all the major aav2 rnas were derived from the minus dna strand and had 3' termini at position 96. the nucleus-specific 4.3- and 3.6-kilobase (kb) rnas had 5' termini at positions 6 and 19, respectively. the 5' term ... | 1980 | 6255215 |
| herpes simplex virus types 1 and 2 completely help adenovirus-associated virus replication. | in addition to adenoviruses, which are capable of completely helping adenovirus-associated virus (aav) multiplication, only herpesviruses are known to provide any aav helper activity, but this activity has been thought to be partial (i.e., aav dna, rna, and protein syntheses are induced, but infectious particles are not assembled). in this study, however, we show that herpes simplex virus type 1 (hsv-1) and type 2 (hsv-2) are in fact complete aav helpers and that aav type 2 (aav2) infectivity yi ... | 1981 | 6270377 |
| mapping of the 5' termini of two adeno-associated virus 2 rnas in the left half of the genome. | the left 45% of the adeno-associated virus 2 genome was sequenced. the 5' termini of two adeno-associated virus-specific rnas were mapped at the nucleotide level within this region of the genome (nucleotides 286 to 288 and 871 to 874). both of these 5' termini map 31 +/- 2 nucleotides downstream from the start of "tata' boxes. | 1982 | 6281463 |
| nucleotide sequence and organization of the adeno-associated virus 2 genome. | the complete nucleotide sequence of the adeno-associated virus 2 genome was determined. the single-stranded genome is 4,675 nucleotides in length and contains inverted terminal repeats of 145 nucleotides, the first 125 nucleotides of which form a palindromic sequence. within the inverted terminal repetitions, there are two distinct sequences representing an inversion of 43 nucleotides that can exist on either terminus. the 5' and 3' termini of three major mrna transcripts, which are present in b ... | 1983 | 6300419 |
| structural proteins of adenovirus-associated virus: subspecies and their relatedness. | capsids of adenovirus-associated virus (aav) are known to contain three major structural proteins (a, b, and c). we have further resolved distinct subspecies of two of the major aav proteins (two forms of protein a and four forms of protein c) which were found in both aav1 and aav2 serotypes. all subspecies were accurately synthesized in a cell-free translation system programmed with rna isolated from infected cells. analysis of virion proteins from the autonomous parvovirus h1 did not reveal a ... | 1983 | 6302317 |
| molecular cloning of adeno-associated virus variant genomes and generation of infectious virus by recombination in mammalian cells. | continued passage of the human parvovirus, adeno-associated virus (aav), at high multiplicity of infection in human cells results in the accumulation of aav particles containing variant genomes. we have analyzed the structure of individual variant aav genomes by molecular cloning in the escherichia coli plasmid, pbr328. each of the aav inserts in six individual recombinant plasmids contained a single internal deletion but in contrast to a previous model, the locations of the deletions were nonra ... | 1984 | 6323482 |
| conformation takes precedence over sequence in adeno-associated virus dna replication. | deletion of an 11-base symmetrical sequence in the inverted terminal repetition of the adeno-associated virus 2 genome inhibits dna replication. substitution of either an 8- or a 12-base symmetrical sequence unrelated to the original 11-base sequence restores dna replication. | 1984 | 6504049 |
| negative regulation of the adeno-associated virus (aav) p5 promoter involves both the p5 rep binding site and the consensus atp-binding motif of the aav rep68 protein. | transcript levels from the p5 promoter of adeno-associated virus type 2 (aav) are negatively regulated by the aav rep78 and rep68 proteins in the absence of helper virus. we have identified a rep-responsive negative cis element of the p5 promoter between the p5 tata box and transcription start site by using 5' and 3' deletions of the p5 promoter fused to the chloramphenicol acetyltransferase gene. this element contains four imperfect gagc repeats similar to the rep recognition sequences (rrss) i ... | 1995 | 7474090 |
| high-level expression of adeno-associated virus (aav) rep78 or rep68 protein is sufficient for infectious-particle formation by a rep-negative aav mutant. | adeno-associated virus (aav) codes for four closely related nonstructural proteins (rep) required for aav dna replication and gene regulation. in vitro studies have revealed that either rep78 or rep68 alone is sufficient for aav dna replication. rep52 and rep40 are not required for dna replication but have been reported to enhance the efficiency of accumulation of single-stranded progeny dna. previous studies on rep-expressing cell lines had indicated that only a subset of the four rep proteins ... | 1995 | 7474103 |
| determination of adeno-associated virus rep68 and rep78 binding sites by random sequence oligonucleotide selection. | to further define the canonical binding site for the p5-promoted rep proteins of the adeno-associated virus, a modified random oligonucleotide selection procedure was performed, using purified recombinant rep protein. these results may explain the effects of rep on cellular gene expression. | 1995 | 7474165 |
| transcriptional transactivation of parvovirus b19 promoters in nonpermissive human cells by adenovirus type 2. | the pathogenic human parvovirus b19 contains a promoter at map unit 6 (b19p6) of the viral genome, expression from which is largely restricted to human cells in the erythroid lineage, whereas a putative promoter at map unit 44 (b19p44) is inactive during a natural viral infection. although nonerythroid human cells, such as hela and kb, allow expression from the b19p6 promoter but not from the b19p44 promoter following dna-mediated transfection, little expression from the b19p6 promoter occurs fo ... | 1995 | 7494329 |
| adeno-associated virus 2-mediated high efficiency gene transfer into immature and mature subsets of hematopoietic progenitor cells in human umbilical cord blood. | recombinant adeno-associated virus 2 (aav) virions were constructed containing a gene for resistance to neomycin (neor), under the control of either the herpesvirus thymidine kinase (tk) gene promoter (vtk-neo), or the human parvovirus b19 p6 promoter (vb19-neo), as well as those containing an upstream erythroid cell-specific enhancer (hs-2) from the locus control region of the human beta-globin gene cluster (vhs2-tk-neo; vhs2-b19-neo). these recombinant virions were used to infect either low de ... | 1994 | 7515101 |
| a maltose-binding protein/adeno-associated virus rep68 fusion protein has dna-rna helicase and atpase activities. | the adeno-associated virus type 2 (aav) rep68 protein produced in escherichia coli as a fusion protein with maltose-binding protein (mbp-rep68 delta) has previously been shown to possess dna-dna helicase activity, as does the purified wild-type rep68. in the present study, we demonstrate that mbp-rep68 delta also catalyzes the unwinding of a dna-rna hybrid. mbp-rep68 delta-mediated dna-rna helicase activity required atp hydrolysis and the presence of mg2+ ions and was inhibited by high ionic str ... | 1995 | 7538173 |
| parvovirus-based vectors for human gene therapy. | it is becoming increasingly clear that the parvovirus-based vectors may prove to be a useful alternative to the more commonly used retroviral vectors in human gene therapy. specifically, the adeno-associated virus 2 (aav), a human parvovirus, has gained particular attention in view of its nonpathogenic nature as well as its remarkable site-specificity of integration into the human chromosome. using the recombinant aav vector system, it is feasible to obtain high-efficiency transduction of slow- ... | 1994 | 7538355 |
| analysis of the complete nucleotide sequences of goose and muscovy duck parvoviruses indicates common ancestral origin with adeno-associated virus 2. | the complete nucleotide sequences of two parvoviruses isolated from goose and muscovy duck were determined. the two virus genomes share 81.9% nucleotide sequence identity, indicating that they are closely related. the coding regions are bracketed by inverted terminal repeats containing palindromes. this is similar to the genome organization of human parvoviruses, adeno-associated virus 2, and b19. amino acid sequence comparison shows that the closest relative of the goose and muscovy duck parvov ... | 1995 | 7571426 |
| adeno-associated virus vectors for gene therapy. | adeno-associated virus type 2 (aav) is a non-pathogenic dna virus which has been utilized as a eukaryotic gene transfer vector in vitro and in vivo. aav possesses a unique set of characteristics which may make it useful for human gene therapy. aav infection does not require host cell proliferation, although expression from aav vectors may exhibit a relative preference for actively dividing cells. both wild-type aav and aav vectors tend to persist in infected cells for prolonged periods of time, ... | 1995 | 7584109 |
| generation of recombinant adeno-associated virus (raav) from an adenoviral vector and functional reconstitution of the nadph-oxidase. | the human parvovirus, adeno-associated virus-2 (aav-2), has many attributes that recommend its use as a gene transfer vehicle, including a broad tissue tropism, the ability to integrate stably into the host genome, and efficient transduction of cells which proliferate slowly. however, application to human gene therapy is currently limited by existing methods for generation of recombinant aav (raav), resulting in relatively low transducing titres. in an attempt to overcome some of these problems, ... | 1995 | 7584126 |
| adeno-associated virus 2-mediated gene transfer and functional expression of the human granulocyte-macrophage colony-stimulating factor. | it is becoming increasingly clear that the adeno-associated virus 2 (aav)-based vector system may prove to be useful for high-efficiency gene transfer in human cells as well as for potential gene therapy in humans. a recombinant aav vector containing the gene for a human hematopoietic growth factor, granulocyte-macrophage colony-stimulating factor (gm-csf), was constructed and used to infect cos-1 cells, a monkey kidney cell line. cos-1 cells infected with the recombinant virus, but not mock-inf ... | 1995 | 7589280 |
| the trans-inhibitory rep78 protein of adeno-associated virus binds to tar region dna of the human immunodeficiency virus type 1 long terminal repeat. | the large rep gene products, rep78 and rep68, of adeno-associated virus (aav) are pleiotropic effector proteins which are required for aav dna replication and the trans-regulation of aav gene expression. apart from these essential functions prerequisite for the life cycle of aav, these rep products are able to inhibit the replication and gene expression of human immunodeficiency virus type 1 (hiv-1) and a number of dna viruses. here, it is demonstrated that rep78, as a chimeric with the maltose ... | 1995 | 7607320 |
| human herpesvirus 6a ts suppresses both transformation by h-ras and transcription by the h-ras and human immunodeficiency virus type 1 promoters. | human herpesvirus 6 strain u1102 (hhv-6a) was shown to contain a 1,473-bp functional transformation suppressor gene (ts). ts exhibited 24% identity and 51% similarity to adeno-associated virus type 2 rep68/78. like adeno-associated virus type 2 rep68/78, hhv-6a ts suppressed h-ras transformation of nih 3t3 cells. suppression of h-ras transformation was eliminated by translation termination linker mutation at amino acid 25, 125, or 245. these data indicated the importance of the c-terminal portio ... | 1995 | 7609062 |
| goose parvovirus--an autonomous member of the dependovirus genus? | goose parvovirus is the etiological agent of derzsy's disease, a fatal hepatitis of young geese. the virus infects geese and muscovy ducks and can be propagated in the laboratory in primary embryonic goose fibroblasts. to date the virus has only been classified by morphological, biochemical, and culture characteristics as an autonomous parvovirus. we now report the cloning and partial sequencing of 3434 nucleotides of the viral genome. three overlapping clones were obtained, encoding regions in ... | 1995 | 7618268 |
| rescue and replication signals of the adeno-associated virus 2 genome. | the adeno-associated virus 2 (aav) genome is a single-stranded dna which contains the inverted terminal repeats (itrs) of 145 nucleotides. the terminal 125 nucleotides of each itr form palindromic hairpin structures that serve as primers for aav dna replication. these hairpin structures also play a crucial role in the integration, as well as the rescue, of the proviral genome from latently-infected cells, or from the recombinant aav plasmids. however, the itrs also contain an additional domain, ... | 1995 | 7623375 |
| intermediates of adeno-associated virus type 2 assembly: identification of soluble complexes containing rep and cap proteins. | the proteins encoded by the adeno-associated virus type 2 (aav-2) rep and cap genes obtained during a productive infection of hela cells with aav-2 and adenovirus type 2 were fractionated according to solubility, cellular localization, and sedimentation properties. the majority of rep and cap proteins accumulated in the nucleus, where they distributed into a soluble and an insoluble fraction. analysis of the soluble nuclear fraction of capsid proteins by sucrose density gradients showed that the ... | 1995 | 7636974 |
| mutational analysis of adeno-associated virus rep protein-mediated inhibition of heterologous and homologous promoters. | the four rep proteins encoded by adeno-associated virus type 2 (aav-2) inhibit transcription of their own promoters and of several heterologous promoters. to gain insight into the molecular mechanism of rep-mediated transcription repression, we studied the effects of the four rep proteins on the accumulation of mrna transcribed from the human papillomavirus type 18 upstream regulatory region hpv18 urr, the human immunodeficiency virus long terminal repeat, and the aav-2 p5 and p19 promoters by t ... | 1995 | 7636994 |
| expression of the cystic fibrosis transmembrane conductance regulator from a novel adeno-associated virus promoter. | adeno-associated virus type 2 (aav) vectors have been used for gene expression in respiratory epithelial cells and may be useful in gene therapy for diseases like cystic fibrosis (cf) which affect the airways. the aav p5 promoter together with the aav inverted terminal repeat (itr) forms a 263-base pair cassette which mediated efficient expression in a cf bronchial epithelial cell line. we report here that the itr itself can mediate gene expression. in stable transfection assays, aav-cf vectors ... | 1993 | 7679117 |
| cloning, expression and purification of full length rep78 of adeno-associated virus as a fusion protein with maltose binding protein in escherichia coli. | the adeno-associated virus (aav) rep78 protein is required for many aspects of aav's life cycle including its dna replication and the regulation of its gene expression. because of increasing utilization of aav as a gene therapy vector and its possible use as an anti-cancer/anti-viral agent, the complete characterization of its rep78 regulatory protein is important. in order to study various functional aspects of rep78, we have cloned and expressed the rep78 gene in escherichia coli using an indu ... | 1995 | 7695627 |
| an improved system for packaging recombinant adeno-associated virus vectors capable of in vivo transduction. | adeno-associated virus (aav) vectors are potentially useful for gene therapy of a number of human diseases. however, the use of these vectors has been limited by the lack of stable vector-packaging cell lines. the difficulties in developing packaging cell lines relate to low levels of rep gene expression from the aav-p5 promoter, and to the propensity of rep proteins to suppress continued growth of immortalized cell lines. we describe here two new techniques which allow these problems to be circ ... | 1995 | 7712331 |
| dissociation of conventional dna binding and endonuclease activities by an adeno-associated virus rep78 mutant. | as part of rep78/68's involvement in adeno-associated virus (aav) dna replication, these highly related, aav encoded proteins bind to the aav terminal repeat (tr) dna and endonucleolytically cleave one strand at the terminal resolution site ("trs" nicking activity) of the tr dna, a site adjacent to the dna binding site, 21 bps from the nearest gctc motif. we have constructed a rep78 mutant, replacing leucine-threonine at amino acids 64-65 with histidine-methionine (64lh65tm). this mutant, expres ... | 1995 | 7763245 |
| sequence elements of the adeno-associated virus rep gene required for suppression of herpes-simplex-virus-induced dna amplification. | herpes simplex virus (hsv) has been shown to induce dna amplification in the host cell genome, which can be suppressed by the adeno-associated virus type 2 (aav-2) rep gene (heilbronn et al., 1990, j. virol. 64, 3012-3018). in an attempt to define domains of rep which are required for this effect a set of expression constructs was generated for rep mutants with either n-terminal and/or c-terminal truncations, with small internal deletions, or with point mutations. in transient cotransfection ass ... | 1995 | 7831779 |
| inhibition of cellular and sv40 dna replication by the adeno-associated virus rep proteins. | in order to define the mechanism used by the adeno-associated virus replication (rep) gene to mediate inhibition of cell proliferation, we have studied its effects on sv40 and cellular dna replication. sv40 dna replication was inhibited by the presence of the rep gene in human 293 cells, and the inhibition was not linked to suppression of sv40 early gene expression. using double-immunofluorescence assays that measured both rep gene expression and bromodeoxyuridine incorporation, we found that th ... | 1995 | 7871734 |
| asymmetric replication in vitro from a human sequence element is dependent on adeno-associated virus rep protein. | the dna of human parvovirus adeno-associated virus type 2 (aav) integrates preferentially into a defined region of human chromosome 19. southern blots of genomic dna from latently infected cell lines revealed that the provirus was not simply inserted into the cellular dna. both the proviral and adjoining cellular dna organization indicated that integration occurred by a complex, coordinated process involving limited dna replication and rearrangements. however, the mechanism for targeted integrat ... | 1995 | 7884849 |
| adeno-associated virus type 2 interferes with early development of mouse embryos. | the human helper-dependent adeno-associated virus type 2 (aav-2) has been shown to induce differentiation in various cell types in culture including pluripotent embryonic cells, in the absence of helper virus. to assess whether induction of differentiation may influence developmental processes we analysed the effect of aav-2 on developing mouse embryos. in vitro infection of fertilized eggs induced arrest of development at the two-cell stage. moreover, micro injection of aav-2 dna (comprising ei ... | 1994 | 7931151 |
| cell lines inducibly expressing the adeno-associated virus (aav) rep gene: requirements for productive replication of rep-negative aav mutants. | the adeno-associated virus (aav) rep gene codes for a family of nonstructural proteins which are required for aav gene regulation and dna replication. in addition, rep has been implicated in a variety of activities outside the aav life cycle which have been difficult to study, since attempts to achieve separate and constitutive expression of rep in stable cell lines have failed so far. here we report the generation of two cell lines which inducibly express rep78 under the control of the glucocor ... | 1994 | 7933099 |
| sequence requirements for stable binding and function of rep68 on the adeno-associated virus type 2 inverted terminal repeats. | replication of the palindromic inverted terminal repeats (itrs) of adeno-associated virus type 2 requires several functions of the viral nonstructural rep proteins. these include binding to the itr, nicking of the double-stranded replication intermediate at the terminal resolution site (trs), and then strand displacement and synthesis from the nick. this report demonstrates the ability of both recombinant fusion maltose-binding protein (mbp)-rep68 delta produced in escherichia coli and wild-type ... | 1994 | 7933128 |
| adeno-associated virus type 2: a latent life cycle. | | 1994 | 7938552 |
| adeno-associated virus vector gene expression occurs in nondividing cells in the absence of vector dna integration. | adeno-associated virus type 2 (aav2)-based vectors are capable of stable expression in the airway epithelium and may be useful for gene therapy for human diseases, such as cystic fibrosis. certain virus vectors, such as retroviruses, require active cell division for integration and expression, but this has not been formally evaluated in the case of aav2. the cystic fibrosis bronchial epithelial cell line, ib3-1, which can be transduced by aav2 vectors, was shown to undergo a decrease in dna synt ... | 1994 | 7946381 |
| the regulatory rep protein of adeno-associated virus binds to sequences within the c-h-ras promoter. | the large rep gene products (rep68 and rep78) of adeno-associated virus (aav) are pleiotropic effector proteins which not only play a critical role in aav dna replication and in the trans-regulation of aav promotor elements, but are also known for their onco-suppressive functions. we have previously demonstrated that the large aav rep protein will strongly inhibit expression from the c-h-ras promoter, but not the murine osteosarcoma virus long terminal repeat (msv-ltr) promoter. to investigate t ... | 1994 | 7954351 |
| mutations in the carboxy terminus of adeno-associated virus 2 capsid proteins affect viral infectivity: lack of an rgd integrin-binding motif. | using site-directed mutagenesis, we tested whether a potential integrin-binding site, (composed of the amino acids rgd) which is predicted in the adeno-associated virus 2 (aav-2) capsid open reading frame (orf), plays a role in the infectivity of aav-2. nucleotide sequencing of wild-type and mutant capsid protein-coding sequences, however, revealed discrepancies with the published sequence data at several positions, including a frameshift in the carboxy terminus which cancels the rgd motif and e ... | 1994 | 7996133 |
| molecular biology of human herpesviruses 6a and 6b. | human herpesvirus 6 variant a (hhv-6a) and human herpesvirus 6 variant b (hhv-6b) are closely related herpesviruses. no disease has been specifically associated with hhv-6a, whereas hhv-6b is the major etiologic agent of exanthem subitum. both viruses may be opportunistic pathogens in the immunocompromised patient. hhv-6 genomes have low g+c contents for herpesviruses (43%); they consist of a 141-kb unique segment that is flanked by single copies of a directly repeated sequence that can vary fro ... | 1993 | 8012736 |
| down-regulation of the human c-fos and c-myc proto-oncogene promoters by adeno-associated virus rep78. | adeno-associated virus (aav) is a non-pathogenic human parvovirus which has anti-tumor and anti-proliferation properties in tissue culture and animal studies. furthermore, aav infection is negatively associated with human cervical cancer. c-myc has been implicated in cervical cancer, and c-fos is involved in signal transduction initiation of cell growth. to study the potential regulation of these two prominent human proto-oncogenes by aav, the expression of three marker coding sequences ligated ... | 1994 | 8012930 |
| characterization of cell lines that inducibly express the adeno-associated virus rep proteins. | the replication (rep) gene of adeno-associated virus (aav) is involved in aav dna replication, gene regulation, and inhibition of cellular transformation induced by various oncogenes. to study the rep gene's antiproliferative effects, we have developed cell lines which express the replication proteins under the control of an inducible mouse metallothionein transcription promoter. the rep78 protein produced in these cell lines binds to the aav terminal repeat sequences in vitro and supports aav d ... | 1994 | 8035483 |
| identification of linear dna sequences that specifically bind the adeno-associated virus rep protein. | we have used baculovirus-expressed rep68 that has been purified to homogeneity to reexamine the binding properties of the rep protein. we find that rep68 is capable of binding to a linear dna sequence that is contained within a 25-bp sequence of the a stem of the adeno-associated virus (aav) terminal repeat proximal to the b and c palindromes. this has been shown conclusively by demonstrating that rep68 could specifically bind to a synthetic oligonucleotide containing the 25-bp region in the abs ... | 1994 | 8035498 |
| interaction of the adeno-associated virus rep protein with a sequence within the a palindrome of the viral terminal repeat. | we have characterized a rep binding sequence which is within the a stem region of the adeno-associated virus terminal repeat (tr) and compared its affinity with that of the complete hairpinned tr for pure rep68. both the a stem and the complete tr substrates produced a complex pattern of protein-dna complexes in which at least six different bound species could be distinguished. competition experiments suggested that the dissociation constant for the a stem sequence is approximately 125-fold high ... | 1994 | 8035499 |
| adeno-associated virus dna replication in vitro: activation by a maltose binding protein/rep 68 fusion protein. | the adeno-associated virus (aav) nonstructural protein rep 68 is required for viral dna replication. an in vitro assay has been developed in which addition of rep 68 to an extract from uninfected hela cells supports aav dna replication. in this paper, we report characterization of the replication process when a fusion of the maltose binding protein and rep 68, expressed in escherichia coli, was used in the assay. replication was observed when the template was either linear double-stranded aav dn ... | 1994 | 8057478 |
| human herpesvirus 6 (hhv-6) is a helper virus for adeno-associated virus type 2 (aav-2) and the aav-2 rep gene homologue in hhv-6 can mediate aav-2 dna replication and regulate gene expression. | we have previously described the apparent acquisition by human herpesvirus 6 (hhv-6) of the multifunctional rep gene of the helper-dependent human parvovirus adeno-associated virus type 2 (aav-2). we report here that hhv-6 is a full helper virus for aav-2 replication, suggesting a mechanism for transfer of the rep gene between the two viruses by recombination of replicative intermediates. the hhv-6 rep gene cloned under control of the human cytomegalovirus immediate early promoter complemented r ... | 1994 | 8091661 |
| intermediates of adeno-associated virus dna replication in vitro. | intermediates of adeno-associated virus type 2 (aav) dna replication in an in vitro assay have been characterized. the assay involves rescue and replication of an aav insert in pbr322. intermediates were shown to be duplex molecules in which at least one terminus was in the extended configuration, in contrast to the hairpinned ends seen after rescue in the absence of aav dna replication. also present were linear double-stranded dimers, which were characterized as either head-to-head or tail-to-t ... | 1994 | 8107263 |
| analysis of adeno-associated virus (aav) wild-type and mutant rep proteins for their abilities to negatively regulate aav p5 and p19 mrna levels. | the rep gene of adeno-associated virus type 2 (aav) encodes four overlapping rep proteins that are involved in gene regulation and replication of the virus. we studied here the regulation of mrna transcribed from the aav p5 and p19 promoters, using transient expression in human 293 cells followed by northern (rna) blot analysis of the mrna. the p5 transcript encodes the larger rep proteins, rep78 and rep68, while the p19 transcript encodes the smaller proteins, rep52 and rep40. a plasmid (pntc3) ... | 1994 | 8151765 |
| cloning, expression, and partial purification of rep78: an adeno-associated virus replication protein. | using the vaccinia virus/t7-rna polymerase transient protein expression system, the aav rep78 protein was expressed in mammalian cells. rep78 protein was found localized primarily to the nucleus of cells. maximal steady-state protein levels were reached as early as 12 hr postinfection, with no discernable increase at later time points. the rep78 protein has been partially purified from nuclear extracts of the expression system. we have successfully used the cloned, purified rep78 protein to comp ... | 1994 | 8178443 |
| site-directed mutagenesis of adeno-associated virus type 2 structural protein initiation codons: effects on regulation of synthesis and biological activity. | it has been shown that two of the three adeno-associated virus type 2 capsid proteins, b and c, are synthesized from a single spliced transcript. protein c arises from an aug codon at nucleotide 2810, whereas protein b is initiated by a unique eucaryotic initiation codon (acg) that lies 65 triplets upstream from the c origin. the third capsid component, protein a, is synthesized from a second spliced transcript which uses an alternative 3' acceptor site. in this study we used oligonucleotide-dir ... | 1994 | 8254726 |
| in vitro replication of adeno-associated virus dna. | the study of eukaryotic viral dna replication in vitro has led to the identification of cellular enzymes involved in dna replication. adeno-associated virus (aav) is distinct from previously reported systems in that it is believed to replicate entirely by leading-strand dna synthesis and requires coinfection with adenovirus to establish completely permissive replication. in previous work, we demonstrated that two of the aav nonstructural proteins, rep78 and -68, are site-specific endonucleases a ... | 1994 | 8289342 |
| adeno-associated virus type 2 rep gene-mediated inhibition of basal gene expression of human immunodeficiency virus type 1 involves its negative regulatory functions. | adeno-associated virus type 2 (aav-2), a human parvovirus which is apathogenic in adults, inhibits replication and gene expression of human immunodeficiency virus type 1 (hiv-1) in human cells. the rep gene of aav-2, which was shown earlier to be sufficient for this negative interference, also down-regulated the expression of heterologous sequences driven by the long terminal repeat (ltr) of hiv-1. this effect was observed in the absence of the hiv-1 transactivator tat, i.e., at basal levels of ... | 1994 | 8289357 |
| biologically active rep proteins of adeno-associated virus type 2 produced as fusion proteins in escherichia coli. | four rep proteins are encoded by the human parvovirus adeno-associated virus type 2 (aav). the two largest proteins, rep68 and rep78, have been shown in vitro to perform several activities related to aav dna replication. the rep78 and rep68 proteins are likely to be involved in the targeted integration of the aav dna into human chromosome 19, and the full characterization of these proteins is important for exploiting this phenomenon for the use of aav as a vector for gene therapy. to obtain suff ... | 1994 | 8289383 |
| suppression of human alpha-globin gene expression mediated by the recombinant adeno-associated virus 2-based antisense vectors. | we sought to investigate the usefulness of the adeno-associated virus 2 (aav)-based vectors to suppress the excess production of the human alpha-globin gene product towards developing a treatment modality for beta-thalassemia since accumulation of free alpha-globin reduces the lifespan of red blood cells in these patients. we constructed recombinant aav virions containing the human alpha-globin gene sequences in antisense orientation driven by the herpesvirus thymidine kinase (tk) promoter, the ... | 1994 | 8294880 |
| identification of a dna-binding domain in the amino terminus of adeno-associated virus rep proteins. | the rep78 and rep68 proteins of adeno-associated virus (aav) bind to the aav terminal repeat hairpin dna and are required for viral replication. we have expressed a series of mutant rep genes from the human immunodeficiency virus type 1 long terminal repeat promoter in human 293 cells and in an in vitro transcription-translation system. mutant proteins were analyzed for aav hairpin dna binding and aav terminal resolution functions. deletion of amino acid residues 523 through 621 of rep 78 had no ... | 1993 | 8380475 |
| versatile adeno-associated virus 2-based vectors for constructing recombinant virions. | we have constructed several plasmid vectors with which a more efficient molecular cloning, followed by rescue, replication, and packaging of dna fragments, can be achieved. the availability of these vectors should facilitate construction of a variety of recombinant adeno-associated virus 2 (aav)-based virions containing therapeutic genes for potential use in human gene therapy. | 1993 | 8383071 |
| analysis of the terminal repeat binding abilities of mutant adeno-associated virus replication proteins. | the adeno-associated virus (aav) rep78 and rep68 proteins play essential roles in viral dna replication, trans activation of viral gene expression, and suppression of oncogene-mediated cellular transformation. by using an extensive set of linker insertion and deletion mutations in the replication gene, we mapped the regions of the rep78 protein that mediate binding to the aav origin of replication in vitro. deletions that removed amino acid codons 25 to 62, 88 to 113, 125 to 256, and 346 to 400 ... | 1993 | 8389942 |
| adeno-associated virus 2-mediated gene transfer in murine hematopoietic progenitor cells. | recombinant human adeno-associated virus 2 (aav) virions were constructed containing a gene for resistance to neomycin (neor), under the control of either the herpesvirus thymidine kinase (tk) promoter (vtk-neo), the murine colony-stimulating factor-1 (csf-1) promoter (vcsf1-neo) or the csf-1 promoter plus an upstream human erythroid cell-specific enhancer, hs-2 (vhs2-csf1-neo). recombinant virions were used to infect low-density murine primary bone marrow cells. in hematopoietic progenitor cell ... | 1993 | 8391471 |