| 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 |
| evidence for covalent attachment of the adeno-associated virus (aav) rep protein to the ends of the aav genome. | we have demonstrated that when the covalently joined ends of linear adeno-associated virus (aav) dna are resolved in vitro, the virus-encoded rep protein becomes covalently attached to the 5' ends of the dna. the covalent bond is between a tyrosine residue of the aav rep protein and a 5' phosphate of a thymidine residue in the aav genome. only the rep protein encoded by the aav p5 promoter, rep68, was capable of becoming covalently attached to the ends of the aav genome; the rep proteins encoded ... | 1990 | 2173787 |
| growth of avian adeno-associated virus in chicken cells transfected with fowl adenovirus serotype 1 dna. | using the chloroquine-modified calcium phosphate coprecipitation technique, fowl adenovirus serotype 1 (fav 1) dna transfects efficiently chicken cell cultures. infection of fav 1 dna transfected cells with helper dependent avian adeno-associated virus (aaav) results in the production of aaav progeny, being detected by an indirect immunofluorescence assay. these findings indicate that fav 1 dna introduced into the host cell promotes actively the growth of aaav. | 1990 | 2176225 |
| conserved region 3 of the adenovirus type 5 dna-binding protein is important for interaction with single-stranded dna. | the adenovirus-encoded single-stranded dna-binding protein (dbp) functions in viral dna replication and several aspects of rna metabolism. previous studies (g. a. m. neale and g. r. kitchingman, j. biol. chem. 264:3153-3159, 1989) have defined three highly conserved regions in the carboxy-terminal domain of the protein (amino acids 178 to 186, 322 to 330, and 464 to 475) that may be involved in the binding of the protein to single-stranded dna. we examined the role of conserved region 3 (464 to ... | 1990 | 2296078 |
| substrate specificity of hela endonuclease r. a g-specific mammalian endonuclease. | we examined the substrate specificity of endonuclease r (endo r) a mammalian endonuclease that cleaves g.c-rich dna sequences. the best substrates for double-stranded cleavage were homopolymeric stretches of poly(dg).poly(dc). plasmids which contain other g-rich sequences were also cleaved but at a reduced frequency. these included the telomeric sequences, d(g4t2) and d(g2-6a), which were cleaved at approximately one-third the frequency of d(g)n.d(c)n. the alternating copolymer d(ga) and the ter ... | 1990 | 2358442 |
| adeno-associated virus rep protein synthesis during productive infection. | adeno-associated virus (aav) rep proteins mediate viral dna replication and can regulate expression from aav genes. we studied the kinetics of synthesis of the four rep proteins, rep78, rep68, rep52, and rep40, during infection of human 293 or kb cells with aav and helper adenovirus by in vivo labeling with [35s]methionine, immunoprecipitation, and immunoblotting analyses. rep78 and rep52 were readily detected concomitantly with detection of viral monomer duplex dna replicating about 10 to 12 h ... | 1989 | 2536109 |
| replication of adeno-associated virus in cells irradiated with uv light at 254 nm. | irradiation of simian virus 40 (ori mutant)-transformed chinese hamster embryo cells (od4 line) with uv light induced a cellular capacity which supported a full cycle of helper-independent adeno-associated virus replication. monochromatic uv light at 254 nm was about 1,000-fold more effective than uv light at 313 nm, indicating that cellular nucleic acid is the primary chromophore in the uv-induced process leading to permissiveness for adeno-associated virus replication. the uv irradiation and t ... | 1989 | 2536816 |
| 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 |
| detection of latent avian adenovirus-associated virus proteins in chicken cells. | chicken kidney cells, derived from the eggs of white leghorn chickens that had serological evidence of prior exposure to both adenovirus and the adeno-associated virus (aav), produced aav antigenic proteins upon challenge with purified adenovirus. antigen was detected by indirect immunofluorescence using monoclonal antibody to aav. the number of fluorescent cells were few and did not increase during the course of adenovirus infection. similar results were obtained using cells prepared from speci ... | 1989 | 2539071 |
| interactions between the termini of adeno-associated virus dna. | the adeno-associated virus (aav) genome is a linear, single polynucleotide chain with inverted terminal repeats of 145 bases. in order to test whether the terminal repeats at opposite ends of the genome have to be able to completely base-pair during dna replication, we have created chimeric genomes in which an 11 base symmetrical sequence has been deleted from the terminal repeat at one end of the genome and replaced by a different 12 base symmetrical sequence. we have used these chimeric constr ... | 1989 | 2539485 |
| 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 |
| factors that bind to adeno-associated virus terminal repeats. | we have identified and characterized a dna-protein complex that forms with the adeno-associated virus (aav) terminal repeats. the complex formed only if the terminal palindrome was in the covalently closed or hairpin configuration; little if any binding was detected with the open duplex form of the terminal repeat. this fact suggested that both secondary structure and primary sequence are essential elements of recognition. dnase i protection studies indicated that virtually all of the a-a' palin ... | 1989 | 2542617 |
| identical ends are not required for the equal encapsidation of plus- and minus-strand parvovirus luiii dna. | sequence analyses of the left and right termini of luiii virus show they are nonidentical imperfect palindromes of 122 and 211 nucleotides, respectively. the left terminus of the minus strand of luiii dna, uniquely in the flip conformation, can assume a t-shaped structure. the right terminus of the minus strand of luiii dna can assume a u-shaped structure, and it exists in either the flip or flop conformation. the termini of luiii shared a high degree of sequence homology and showed conserved se ... | 1989 | 2542625 |
| organization of adeno-associated virus dna in latently infected detroit 6 cells. | the dna of a human dependovirus, adeno-associated virus (aav), integrates into the cellular genome under conditions nonpermissive for viral dna replication. the aav dna can be rescued from the cellular genome by superinfection with a helper virus, e.g., adenovirus. to characterize the organization of the proviral dna in greater detail, we isolated three proviral subfragments from a latently infected human cell line. our findings, based on sequence analysis, restriction enzyme mapping, and genomi ... | 1989 | 2543124 |
| replication of a human parvovirus nonsense mutant in mammalian cells containing an inducible amber suppressor. | when recombinant plasmids containing the entire adeno-associated virus (aav) genome are transfected into permissive cells infected with a helper adenovirus, infectious aav particles are efficiently generated. these plasmids can be used to generate mutant aav genomes or recombinant aav vectors. packaging of mutant aav genomes has required complementation with a second aav plasmid in the transfection assay which may lead to generation of significant amounts of wild-type aav recombinants. one appro ... | 1989 | 2545030 |
| adeno-associated virus p5 promoter contains an adenovirus e1a-inducible element and a binding site for the major late transcription factor. | activity of the adeno-associated virus p5 transcriptional control region was found to be induced by adenovirus e1a gene products. a pair of adjacent sequence elements was found to mediate both basal and e1a-induced p5 activity. the first element is a binding site for the major late transcription factor (mltf), a factor first identified on the basis of its binding to a specific sequence within the adenovirus major late promoter. the second element is a tandemly repeated 10-base-pair sequence whos ... | 1989 | 2545917 |
| dna amplification in genetic toxicology. | chemical compounds can cause amplification of specific dna sequences. dna amplification may result in an enhanced production of gene products which help cells to cope with the chemicals. this may lead to a resistance of the cells toward the agent. additionally, initiation of transformation or progression of transformed cells to tumorigenicity may also involve dna amplification. therefore, it is of interest to study the potential of chemicals to induce dna amplification. this report focuses on th ... | 1989 | 2546072 |
| helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression. | a method is described for the production of recombinant adeno-associated virus (aav) stocks that contain no detectable wild-type helper aav. the recombinant viruses contained only the terminal 191 nucleotides of the aav chromosome bracketing a nonviral marker gene. trans-acting aav functions were provided by a helper dna in which the terminal 191 nucleotides of the aav chromosome were substituted with adenovirus terminal sequences. although the helper dna did not appear to replicate, it expresse ... | 1989 | 2547998 |
| 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 |
| amplification of bovine papillomavirus dna by n-methyl-n'-nitro-n-nitrosoguanidine, ultraviolet irradiation, or infection with herpes simplex virus. | treatment with n-methyl-n'-nitro-n-nitrosoguanidine (mnng) or irradiation with ultraviolet light (uv254 nm) induces amplification of integrated as well as episomal sequences of bovine papillomavirus (bpv) type 1 dna in bpv-1-transformed mouse c127 cells (i.e., id13 cells). this is shown by filter in situ hybridization and southern blot analysis of cellular dna. similarly, infection of id13 cells with herpes simplex virus (hsv) type 1 which has been shown to be mutagenic for host cell dna leads t ... | 1989 | 2549724 |
| expression from the adeno-associated virus p5 and p19 promoters is negatively regulated in trans by the rep protein. | the leftward two promoters of the adeno-associated virus (aav) 2 genome were fused to reporter genes, and the constructs were used to transfect hela cells. the promoters functioned constitutively but were repressed in trans by the aav rep gene product(s). the repression was relieved by adenovirus infection. evidence which indicated an enhancer function for the inverted terminal repeat of the aav-2 genome was also obtained. | 1989 | 2550677 |
| 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 |
| mutagenesis of an aug codon in the adeno-associated virus rep gene: effects on viral dna replication. | the adeno-associated virus (aav) rep gene is transcribed from two promoters, p5 and p19, which code for two over-lapping families of rep proteins. the proteins coded by p5 transcripts contain an amino-terminal domain not present in the proteins coded by p19 transcripts. the rep gene is required for aav dna replication and also mediates pleiotropic effects in positive and negative regulation of expression of genes driven by either aav or heterologous promoters. all three functions require rep pro ... | 1989 | 2554565 |
| biochemical analysis of adenovirus type 5 dna-binding protein mutants. | we previously reported the isolation and functional characterization of seven adenovirus type 5 (ad5) dna-binding protein (dbp) point mutants (quinn, c. o., and kitchingman, g. r. (1986) j. virol. 60, 653-661). six of the seven mutants were defective in their ability to help adeno-associated virus replicate its dna. to determine the level at which the mutations affect this function of the dbp, we analyzed several properties of the mutant proteins. all are transported to the nucleus and are post- ... | 1989 | 2644280 |
| adenovirus early region 4 encodes two gene products with redundant effects in lytic infection. | in order to assign specific functions to individual gene products encoded by adenovirus type 5 early region 4 (e4), we have constructed and analyzed a set of mutant viruses that express individual e4 open reading frames or combinations of open reading frames. the results of these analyses demonstrate that the gene products of e4 open reading frames 3 and 6 have redundant effects in viral lytic infection. these e4 products independently augment viral dna replication, viral late protein synthesis, ... | 1989 | 2724411 |
| gene expression in adeno-associated virus vectors: the effects of chimeric mrna structure, helper virus, and adenovirus va1 rna. | we used a recombinant plasmid containing an adeno-associated virus (aav) genome to construct several vectors which express the gene for chloramphenicol acetyltransferase (cat). we transfected four different aav-cat vectors into human 293 (adenovirus-transformed) cells and analyzed cat activity. we show that, for vectors using the aav p40 and p19 promoter, the chimeric aav-cat transcripts began from the correct 5' position but the basal level of cat expression depended in part on the structure of ... | 1987 | 2820138 |
| replication of the adeno-associated virus dna termini in vitro. | the validity of the model proposed by cavalier-smith for the replication of linear, single-stranded dna molecules was tested by using subgenomic dna termini isolated from adeno-associated virus (aav), a defective parvovirus. in purified dna replication systems, the terminal hairpin structure of aav dna with a 3'-oh end was shown to prime the synthesis of the daughter dna strand, which was covalently linked to the parental strand. this resulted in the synthesis of a duplex dna structure with a te ... | 1987 | 2822604 |
| characterization of adeno-associated virus rep proteins in human cells by antibodies raised against rep expressed in escherichia coli. | the rep gene of the defective human parvovirus, adeno-associated virus, (aav) mediates several trans-acting functions important to virus replication, transcription, and gene expression. at least four overlapping polypeptides are expressed from the rep gene. we have constructed a prokaryotic vector which expressed in escherichia coli a region of aav comprising 93% of the largest aav rep protein. the protein expressed in e. coli, rep 78.93, was used to raise specific antibodies in rabbits. these a ... | 1987 | 2823460 |
| adenovirus e1b 55-mr polypeptide facilitates timely cytoplasmic accumulation of adeno-associated virus mrnas. | adenovirus provides helper functions that facilitate replication of adeno-associated virus (aav). both the adenovirus e1b 55-mr and e4 34-mr polypeptides are required for efficient and timely accumulation of aav mrna, proteins, and dna. the e1b 55-mr polypeptide is also required for rescue of the integrated aav genome in detroit 6-d5 cells in a normal time frame. all of these effects probably result from a single, primary delay in aav mrna accumulation. the aav helper function provided by the e1 ... | 1988 | 2824848 |
| regulation of adeno-associated virus gene expression in 293 cells: control of mrna abundance and translation. | we studied the effects of the adeno-associated virus (aav) rep gene on the control of gene expression from the aav p40 promoter in 293 cells in the absence of an adenovirus coinfection. aav vectors containing the chloramphenicol acetyltransferase (cat) gene were used to measure the levels of cat expression and steady-state mrna from p40. when the rep gene was present in cis or in trans, cat expression from p40 was decreased 3- to 10-fold, but there was a 2- to 4-fold increase in the level of p40 ... | 1988 | 2824856 |
| gene transfer into hematopoietic progenitor cells mediated by an adeno-associated virus vector. | we describe here the transduction of murine hematopoietic progenitor cells with the dominant selectable neomycin drug-resistance (neo) gene using a recombinant adeno-associated virus (aav) vector. successful transformation of progenitor cells to drug resistance was determined to be approximately 1.5% by colony formation in the presence of geneticin sulfate (g-418). the value of aav as an alternative to the retrovirus vector systems is discussed. | 1988 | 2829430 |
| avian adeno-associated parvovirus and marek's disease virus: studies of viral interactions in chicken embryo fibroblasts. brief report. | using growth kinetics we demonstrate two effects based on interactions between the chicken herpesvirus, marek's disease virus (mdv), and the dependovirus, avian adeno-associated virus (aaav), in coinfected chicken embryo fibroblasts (cef): (i) mdv provides helper activity for an efficient multiplication of aaav; (ii) a high multiplicity of coinfecting aaav inhibits completely the growth of mdv as well as aaav. | 1988 | 2831859 |
| the adeno-associated virus rep gene inhibits replication of an adeno-associated virus/simian virus 40 hybrid genome in cos-7 cells. | a hybrid adeno-associated virus (aav)/simian virus 40 (sv40) genome is described. in this construct sv40 regulatory sequences, including the early promoter/enhancers and origin of dna replication, were substituted for the aav p5 promoter, which normally controls expression of the aav rep gene. the hybrid genome was phenotypically indistinguishable from wild-type aav in human cells in the presence or absence of helper virus. upon transfection into cos-7 cells, which constitutively produced the sv ... | 1988 | 2833621 |
| dna amplification of adeno-associated virus as a response to cellular genotoxic stress. | we studied dna amplification of helper virus-dependent parvoviruses [adeno-associated virus (aav)] following genotoxic treatment of a number of mammalian cell lines from different species including primary, immortalized, and tumorigenic cells. all cell lines, either infected with aav or transfected with parvoviral dna, readily amplified aav dna in the absence of helper virus following treatment of cells with a wide variety of genotoxic agents like chemical carcinogens, uv, heat shock, and metabo ... | 1988 | 2835153 |
| adeno-associated virus general transduction vectors: analysis of proviral structures. | we used two kinds of adeno-associated virus (aav) vectors to transduce the neomycin resistance gene into human cells. the first of these (dl52-91) retains the aav rep genes; the second (dl3-94) retains only the aav terminal repeats and the aav polyadenylation signal (428 base pairs). both vectors could be packaged into aav virions and produced proviral structures that were essentially the same. thus, the aav sequences that are required in cis for packaging (pac), integration (int), rescue (res), ... | 1988 | 2835501 |
| 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 |
| cellular transcription factor binds to adenovirus early region promoters and to a cyclic amp response element. | we have analyzed the sequences that regulate the transcription of adenovirus type 5 early region 4 (e4). a region located immediately upstream of the e4 tata box is required for efficient e4 transcription in vitro and in vivo. a cellular transcription factor, termed etf-a, binds to this region. etf-a also binds to additional sites in the e4 5'-flanking region, including the adenovirus terminal repeat, as well as to the adenovirus early region 2 promoter region and the adeno-associated virus earl ... | 1988 | 2839717 |
| alternate mrna splicing is required for synthesis of adeno-associated virus vp1 capsid protein. | fine-structure mapping of the capsid-specific mrnas from adeno-associated virus (aav) revealed an alternate splicing pattern in these rnas. s1 nuclease and primer extension analyses showed that splicing of these mrnas occurs at acceptor sites at nucleotide 2228 (major splice) or 2201 (minor splice). both splice acceptors were ligated to the same 55-nucleotide leader in mature mrnas. both species were present in equal amounts in mrna derived from aav plasmid-transfected cells. however, when adeno ... | 1988 | 2841488 |
| in vitro excision of adeno-associated virus dna from recombinant plasmids: isolation of an enzyme fraction from hela cells that cleaves dna at poly(g) sequences. | when circular recombinant plasmids containing adeno-associated virus (aav) dna sequences are transfected into human cells, the aav provirus is rescued. using these circular aav plasmids as substrates, we isolated an enzyme fraction from hela cell nuclear extracts that excises intact aav dna in vitro from vector dna and produces linear dna products. the recognition signal for the enzyme is a polypurine-polypyrimidine sequence which is at least 9 residues long and rich in g.c base pairs. such sequ ... | 1988 | 2841582 |
| sequence and symmetry requirements within the internal palindromic sequences of the adeno-associated virus terminal repeat. | the defective parvovirus, adeno-associated virus (aav), contains a single-stranded dna genome of 4681 bases with inverted terminal repeats of 145 bases. the distal 125 bases of the repeat are palindromic allowing a hairpin to form for initiation of dna synthesis. the palindromic region contains three palindromes, two smaller internal palindromes flanked by a larger palindrome, which allow the hairpinned dna to assume a t-shaped conformation during dna replication. deletion of an internal palindr ... | 1988 | 2845646 |
| effect of a viral rep gene on transformation of cells by an adeno-associated virus vector. | adeno-associated virus (aav) vectors readily express the gene for geneticin-resistance under control of the aav p40 promoter when chromosomally integrated at low copy number in mammalian cells. we show that a truncated aav rep gene, transcribed from the p5 and p19 promoters, mediates a negative effect on expression of geneticin-resistance in human 293 cells and a positive effect in hela cells. also, we describe a novel phenotype for a mutant expressing the p19 rep gene alone which has a negative ... | 1988 | 2845666 |
| adeno-associated virus: a vector system for efficient introduction and integration of dna into a variety of mammalian cell types. | adeno-associated virus (aav) is a single-stranded dna parvovirus that is dependent on adenovirus or herpesvirus for reproductive functions. we describe the construction of recombinant aav vectors containing the chloramphenicol acetyltransferase gene or the neomycin phosphotransferase gene. these vectors carried their respective genes into a wide variety of cell types, including primary skin fibroblasts and hematopoietic cells. infection efficiencies varied with cell type and ranged up to 3.0%. c ... | 1988 | 2847025 |
| organization of the adeno-associated virus (aav) capsid gene: mapping of a minor spliced mrna coding for virus capsid protein 1. | we have mapped a minor spliced transcript with mrna properties which is derived from the capsid gene promoter (p40) of the adeno-associated virus (aav) genome. by s1 nuclease mapping as well as primer extension analysis this mrna was found to occur by splicing at the same donor site (nucleotide, nt 1907) but at an alternative acceptor site when compared to the major, 2.3-kb spliced p40 transcript which encodes two of the three aav capsid proteins, namely vp2 and vp3. this minor acceptor site is ... | 1988 | 2847413 |
| perturbation of the cell cycle by adeno-associated virus. | infection of nonpermissive cells with adeno-associated virus (aav) or aav inactivated by uv light inhibited their multiplication in culture and interfered with their transit through the cell cycle. the perturbation of the cell cycle led to the accumulation of cells in the late s and/or g2 phases. the aav-mediated inhibition of growth was dependent upon high concentrations of input virus and the types of cells. presenescent embryonic fibroblasts of syrian hamster and human origin were the most se ... | 1988 | 2849233 |
| regulation of adeno-associated virus dna replication. | | 1988 | 2850017 |
| purification method for the avian adeno-associated virus. | avian adeno-associated virus (aaav) pre-purified by uvasol extraction, one discontinuous and two cscl equilibrium density gradients, was still considerably contaminated with avian adenovirus (celo strain). four different approaches were investigated in attempts to improve the elimination of the contaminating celo virus. the contaminations were assayed by double immunodiffusion and indirect immunofluorescence. the immunoprecipitation of celo virus with antiserum and protein a-sepharose is the mos ... | 1985 | 2989313 |
| comparison of the dna sequence and secondary structure of the herpes simplex virus l/s junction and the adeno-associated virus terminal repeat. | the defective parvovirus adeno-associated virus (aav) is absolutely dependent upon coinfection with either adenovirus or herpes simplex virus (hsv) for its multiplication. we have compared the terminal repeats of hsv-1f strain dna with the terminal 200 nucleotides of aav dna. our findings demonstrate similarities between portions of the hsv inverted repeats found at the l/s junction and the termini of aav. by computer analysis we have determined potential secondary folding patterns for both geno ... | 1985 | 2995732 |
| 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 |
| human cytomegalovirus completely helps adeno-associated virus replication. | coinfection of adeno-associated virus (aav) with human cytomegalovirus (hcmv) strain towne in human embryonic fibroblasts resulted in accumulation of aav capsid antigen and production of infectious aav with a lag of 24 hr compared to aav replication in aav-adenovirus coinfections. in contrast to previous observation, these findings demonstrated that hcmv is a competent helper virus for the complete replication of aav. in addition, hcmv and aav were synergistic in their cytopathic effects on cell ... | 1985 | 2998066 |
| 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 |
| herpesviruses provide helper functions for avian adeno-associated parvovirus. | the avian herpesviruses infectious laryngotracheitis virus (iltv) and herpesvirus of turkeys (hvt), as well as the mammalian herpesvirus pseudorabies virus (prv) were able to provide complete helper activity for the production of infectious avian adeno-associated virus (aaav) in chicken cells. the presence of aaav in the infected chicken cell reduced the multiplication of hvt. iltv or prv, however, were not affected if used as helper viruses. infectious aaav was determined by an indirect immunof ... | 1986 | 3003233 |
| effect of adeno-associated virus on transformation of nih 3t3 cells by ras gene and on tumorigenicity of an nih 3t3 transformed cell line. | transfection of nih-3t3 cells with the plasmid pj234, containing dna from the human bladder carcinoma t24 cell line (ras gene), results in their transformation. adeno-associated virus did not affect significantly the number of the transformed foci when different multiplicities of infection were used and when the virus was added to the cultures at different time intervals before or after transfection. a transformed cell line was derived following transfection of nih 3t3 cells by the ras gene. inf ... | 1986 | 3009007 |
| evolution of a defective virus from a cellular defense mechanism. | adeno-associated virus is a defective dna virus, requiring the presence of a helper virus in order to replicate. in this paper we consider its origin in light of several observations, most notably the following: its own replication inhibits that of the helper virus; its dna structure resembles that of transposable (moveable) elements; and extrachromosomal circles of dna, about the size of adeno-associated virus dna, have been found recently in eukaryotic cells. we have arrived at a hypothesis co ... | 1986 | 3012208 |
| vaccinia virus, herpes simplex virus, and carcinogens induce dna amplification in a human cell line and support replication of a helpervirus dependent parvovirus. | the sv40-transformed human kidney cell line, nb-e, amplifies integrated as well as episomal sv40 dna upon treatment with chemical (dmba) or physical (uv irradiation) carcinogens ("initiators") as well as after infection with herpes simplex virus (hsv) type 1 or with vaccinia virus. in addition it is shown that vaccinia virus induces sv40 dna amplification also in the sv40-transformed chinese hamster embryo cell line, co631. these findings demonstrate that human cells similar to chinese hamster c ... | 1986 | 3012864 |
| 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 |
| adeno-associated virus vector for high-frequency integration, expression, and rescue of genes in mammalian cells. | we describe the construction of an adeno-associated virus (aav) vector in which the coding sequence of the procaryotic gene neo is expressed under the control of the major aav promoter p40. this aav-neo vector allowed stable expression of neo as a dominant selective marker in mammalian cells by selection of cells which were resistant to the antibiotic geneticin (g418). when the vector was introduced into human (293 or hela) cell lines by a dna transfection procedure, stable geneticin-resistant c ... | 1985 | 3018511 |
| 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 |
| functional analysis of the adenovirus type 5 dna-binding protein: site-directed mutants which are defective for adeno-associated virus helper activity. | we generated four point mutations in the dna-binding protein (dbp) gene of adenovirus type 5 by oligonucleotide-directed site-specific mutagenesis. the sites mutated were in the three conserved regions (cr; amino acids 178-186 [cr1], 322-330 [cr2], and 464-475 [cr3]) identified previously by comparative sequence analysis (g. r. kitchingman, virology 146:90-101, 1985). the mutations resulted in changes in amino acids 181 (trp to leu), 323 (arg to leu), 324 (trp to leu), and 469 (phe to ile). the ... | 1986 | 3021998 |
| identification of the trans-acting rep proteins of adeno-associated virus by antibodies to a synthetic oligopeptide. | prior genetic analysis provided evidence for trans-acting regulatory proteins (rep) coded by the left-hand open reading frame (orf-1) of adeno-associated virus (aav). we have used immunoblotting analysis to identify four protein products of orf-1. antibodies elicited against an oligopeptide encoded by orf-1 were reacted with extracts of cells that were infected with aav or transfected with aav recombinant vectors in the presence or absence of helper adenovirus. the antibody recognized four polyp ... | 1986 | 3023672 |
| replication of adeno-associated virus in synchronized cells without the addition of a helper virus. | we investigated the helper-independent replication of adeno-associated virus (aav) in cells synchronized by pretreatment with hydroxyurea, reversal of polyamine depletion, or physical mitotic detachment. in chinese hamster cells (od4 line) treated with hydroxyurea prior to infection. aav underwent a complete cycle of replication. transfection of such cells with plasmid-cloned aav dnas also gave rise to infectious viral progeny. synchronization of od4 cells by reversal of polyamine depletion or m ... | 1987 | 3029431 |
| adeno-associated virus gene expression inhibits cellular transformation by heterologous genes. | in this paper we report that adeno-associated virus (aav) genomes inhibit stable transformation by several dominant selectable marker genes upon cotransfection into mouse tissue culture cells. cotransfection of aav genomes also inhibited the expression of psv2cat in transient assays. in both cases, the inhibitory effect was independent of aav dna replication but required the aav p5 and p19 genes, which encode proteins required for aav dna replication and regulation of aav gene expression. finall ... | 1987 | 3037312 |
| a recombinant plasmid from which an infectious adeno-associated virus genome can be excised in vitro and its use to study viral replication. | a recombinant plasmid carrying an infectious adeno-associated viral genome was constructed that differs in several key respects from previously described recombinants. first, the vector is pembl8(+), which allows isolation of viral plus and minus strands. second, the inserted viral sequences contain two xbai cleavage sites that flank the viral coding domain. these inserts do not affect replication of the virus, and they allow nonviral sequences to be easily inserted between the cis-acting termin ... | 1987 | 3041032 |
| cloning of the human thyrotropin beta-subunit gene and transient expression of biologically active human thyrotropin after gene transfection. | a 17 kilobase pair fragment of dna containing the human tsh (htsh) beta-subunit gene was isolated from a human leukocyte genomic library. using a 621 base pair human cg alpha-subunit cdna and a 2.0 kilobase pair genomic fragment of htsh beta containing both coding exons, we constructed hcg alpha and htsh beta expression vectors containing either the early promoter of simian virus 40 or the promoters of adeno-associated virus. cotransfection of two adeno-associated virus vectors, each containing ... | 1988 | 3398841 |
| negative and positive regulation in trans of gene expression from adeno-associated virus vectors in mammalian cells by a viral rep gene product. | we previously described use of the human parvovirus, adeno-associated virus (aav), as a vector for transient expression in mammalian cells of the gene for chloramphenicol acetyltransferase (cat). in the aav vector, pts1, the cat gene is expressed under the control of the major aav promoter p40. this promoter is embedded within the carboxyl-terminal region of an open reading frame (orf-1) which codes for a protein (rep) required for aav dna replication. we show here that the rep product has addit ... | 1986 | 3491293 |
| novel transcription map for the b19 (human) pathogenic parvovirus. | the b19 parvovirus, a small single-stranded dna virus of 5.4 kilobases, is pathogenic in humans. b19 has remarkable specificity for erythroid progenitor cells and has been propagated in vitro only with human erythroid bone marrow. replication of viral dna and the viral protein products of b19 appear similar to those of other animal parvoviruses. however, b19 transcription had unusual features in comparison with that in other animal parvoviruses. at least nine overlapping poly(a)+ transcripts wer ... | 1987 | 3599180 |
| [an adeno-associated virus vector]. | | 1987 | 3672936 |
| electron microscopy of dna from adeno-associated virus type i. | | 1971 | 4101807 |
| studies on the relationship between adeno-associated virus type i (aav-1) and adenoviruses. i. replication of aav-1 in certain cell cultures and its effect on helper adenovirus. | | 1967 | 4290509 |
| oncogenicity of mixtures of adeno-associated virus and adenovirus type 12. | | 1968 | 4302692 |
| replication of an adeno-associated virus in canine and human cells with infectious canine hepatitis virus as a "helper". | | 1967 | 4318961 |
| detection and localization of viruses in human fetal intestinal organ cultures by immunofluorescence. | viral antigens from viruses belonging to four different viral groups were detected directly in human fetal intestinal organ cultures by the application of immunofluorescent techniques. the time of appearance and the cellular localization of fluorescent-stainable antigen varied with the type of virus under investigation. after infection with adenovirus or with adeno-associated virus, fluorescent-stainable antigen was seen in the epithelial cells of the explants, though no light microscopic change ... | 1972 | 4344634 |
| arrangement of nucleotide sequences in adeno-associated virus dna. | | 1973 | 4586409 |
| letter: visualization of the inverted terminal repetition in adeno-associated virus dna. | | 1974 | 4816655 |
| studies on the relationship between adeno-associated virus type 1 (aav-1) and adenoviruses. ii. inhibition of adenovirus plaques by aav; its nature and specificity. | | 1967 | 4964865 |
| the potentiation of type 1 adeno-associated virus by temperature-sensitive conditional-lethal mutants of celo virus at the restrictive temperature. | | 1971 | 5000131 |
| separation of two types of adeno-associated virus particles containing complementary polynucleotide chains. | an adeno-associated virus containing bromodeoxyuridine-substituted deoxyribonucleic acid has been fractionated by equilibrium centrifugation in cscl into two classes of virions which contain complementary polynucleotide chains. | 1972 | 5014934 |
| nucleic acid from an adeno-associated virus: chemical and physical studies. | | 1966 | 5229859 |
| haemagglutinating adeno-associated virus (aav) in association with bovine adenovirus type 1. brief report. | | 1970 | 5530958 |
| oncogenicity of mixtures of adeno-associated virus and adenovirus type 12. | | 1968 | 5659627 |
| inhibition of adenovirus 12 oncogenicity by adeno-associated virus. | | 1968 | 5668107 |
| 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 |
| requirement for either early region 1a or early region 1b adenovirus gene products in the helper effect for adeno-associated virus. | several adenovirus early genes act together to promote growth of the helper-dependent adeno-associated virus (aav). data from several laboratories have implicated adenovirus early regions 1a, 1b, 2a, and 4 in the helper effect, as well as the small rna polymerase iii transcript, virus-associated rna i. although a subset of these must participate directly in the aav life cycle, some may play an indirect role by influencing expression of the others. this paper is concerned particularly with the ro ... | 1984 | 6086952 |
| rescue of adeno-associated virus from recombinant plasmids: gene correction within the terminal repeats of aav. | we have isolated three types of pbr322-aav recombinant plasmids that contain deletions within the 145 bp aav terminal repeats. when the plasmids were transfected into human cells, mutants that contained deletions within the left (type i) or right (type ii) terminal repeat were viable. of four mutants examined that contained deletions in both termini (type iii), only one was viable. all of the viable mutants produced aav virions that contained wild-type aav dna. furthermore, the viable type iii d ... | 1983 | 6088052 |
| genetic analysis of adeno-associated virus: properties of deletion mutants constructed in vitro and evidence for an adeno-associated virus replication function. | transfection of a pbr322-based, recombinant plasmid, pav2, containing the entire adeno-associated virus (aav) type 2 genome into human 293 cells in the presence of helper adenovirus resulted in rescue and replication of aav to yield infectious particles. we constructed mutants of pav2 containing deletions within the aav sequence. we describe here the phenotypes of these aav deletion mutants. the results can be summarized as follows. mutants (cap-) with deletions between map positions 53 and 85 d ... | 1984 | 6088786 |
| 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 |
| use of adeno-associated virus as a mammalian dna cloning vector: transduction of neomycin resistance into mammalian tissue culture cells. | adeno-associated virus (aav) is a human dna virus that has a broad host range and can be grown both as an integrated provirus and as a lytic genome. these properties suggested that aav may be useful as a mammalian transduction vector. to test this possibility, we have isolated a recombinant aav viral stock in which the neomycin resistance gene was substituted for the aav capsid genes. using this recombinant stock, we have demonstrated that aav can be used to transduce foreign dna into human and ... | 1984 | 6093102 |
| replication of adeno-associated virus dna. complementation of naturally occurring rep- mutants by a wild-type genome or an ori- mutant and correction of terminal palindrome deletions. | when the entire adeno-associated virus (aav) genome is inserted into a bacterial plasmid, infectious aav genomes can be rescued and replicated when the recombinant aav-plasmid dna is transfected into human 293 cells together with helper adenovirus particles. we have taken advantage of this experimental system to analyze the effects of several classes of mutations on replication of aav dna. we obtained aav mutants by molecular cloning in bacterial plasmids of naturally occurring aav variant or de ... | 1984 | 6094825 |
| a human parvovirus, adeno-associated virus, as a eucaryotic vector: transient expression and encapsidation of the procaryotic gene for chloramphenicol acetyltransferase. | we have used the defective human parvovirus adeno-associated virus (aav) as a novel eucaryotic vector (parvector) for the expression of a foreign gene in human cells. the recombinant, pav2, contains the aav genome in a pbr322-derived bacterial plasmid. when pav2 is transfected into human cells together with helper adenovirus particles, the aav genome is rescued from the recombinant plasmid and replicated to produce infectious aav particles at high efficiency. to create a vector, we inserted a pr ... | 1984 | 6095038 |
| the genetics of adeno-associated virus. | | 1984 | 6098150 |
| adeno-associated virus rna transcription in vivo. | we have studied rna transcripts of the defective parvovirus, adeno-associated virus (aav) present in poly(a)rich and poly-(a)free fractions of nuclear and cytoplasmic rna prepared from cells infected together with a helper adenovirus. cytoplasmic poly-(a)rich rna contains three overlapping spliced aav rnas having sizes of 3.9 x 10(3), 3.3 x 10(3) and 2.3 x 10(3) bases respectively. the nuclear precursors of these rnas appear to be the coterminal unspliced poly(a)-rich rnas containing 4.2 x 10(3) ... | 1981 | 6173214 |
| virological studies in amyotrophic lateral sclerosis. | complement-fixing antibody response to eleven different viruses were measured in amyotrophic lateral sclerosis (als) patients, contacts of als patients, neurological controls, and normal controls. the normal controls showed a decreased response to adeno-associated virus and an increased response to adenovirus when compared to the other groups. levels of interferon-like substances also were investigated in sera and cerebrospinal fluids of als patients and neurological controls. responses were of ... | 1982 | 6175901 |
| analysis of proteins, helper dependence, and seroepidemiology of a new human parvovirus. | a new type of defective parvovirus, tentatively designated as adeno-associated virus type 5 (aav-5), is characterized as far as its proteins, its helper dependence, and its seroepidemiology are concerned. the protein analysis of aav-5 in polyacrylamide gels demonstrated the presence of three structural polypeptides, corresponding to vp 1, vp 2, and vp 3 of other aav types. the preparation of monoclonal antibodies against aav-5 permitted the analysis of viral structural antigen expression by usin ... | 1984 | 6200995 |
| molecular structure of adeno-associated virus variant dna. | when lysates of human cells, infected jointly with the defective parvovirus, adeno-associated virus (aav), and a helper adenovirus, are banded to equilibrium in cscl buoyant density gradients, virus particles of various densities are obtained. infectious aav particles mainly band at a density of 1.41 g/cm3 with a minor component at 1.45 g/cm3. noninfectious aav particles band at densities between 1.41 and 1.32 g/cm3. we have analyzed, by mapping with site-specific endodeoxyribonucleases, the mol ... | 1980 | 6244311 |
| heavy and light particles of adeno-associated virus. | kb cells coinfected with adenovirus and adeno-associated virus (aav) yielded two kinds of infectious aav particles that banded in cscl at densities of 1.45 and 1.41 g/cm(2), respectively. the 1.45 band was found to be composed of a heterogeneous group of viral particles that could be subfractionated by velocity sedimentation. the main component from this band had a smaller s value (109) than the main component from the 1.41 band (111s), although both had the same dna/protein ratio and the same d ... | 1980 | 6245263 |
| assembly of adeno-associated virus. | | 1980 | 6245509 |
| 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 |
| adenovirus early region 1b gene function required for rescue of latent adeno-associated virus. | | 1980 | 6249041 |
| integration of the adeno-associated virus genome into cellular dna in latently infected human detroit 6 cells. | a clone of human cells (detroit 6) latently infected by adeno-associated virus (aav) has been characterized with regard to the status of the viral dna. in both early (9 to 10) and late (118) passages of the clone, aav-dna was recombined with host dna, at least in some cases as a head-to-tail tandem repeat, via the terminal sequences of the viral genome. however, it was not possible to distinguish between integration into chromosomal dna and very large plasmids (< 20 x 10(6) molecular weight) whi ... | 1980 | 6251245 |
| adenovirus helper function for growth of adeno-associated virus: effect of temperature-sensitive mutations in adenovirus early gene region 2. | adeno-associated virus (aav) grows efficiently only in cells that are also infected with an adenovirus (ad). we employed ad mutants to determine which genes may be required for the aav helper function. two mutants of ad type 5 (ad5), ad5ts125 and ad5ts107, with temperature-sensitive lesions in the e72 dna-binding protein coded by the ad early region 2, were deficient for aav helper functions at the nonpermissive temperature (40 degrees c). in contrast, ad5ts149, with a temperature-sensitive lesi ... | 1980 | 6251278 |