| hamster embryo cells transformed by herpes simplex virus: reactions with adeno-associated satellite virus (aav) and its adenovirus helpers. | we have studied the reactions of hamster embryo cells transformed by ultraviolet-inactivated herpes simplex type 2 (333-8-9 t cells) to infections with adeno-associated satellite virus (aav) and its adenovirus helpers. resident hsv structural antigens were not detectable in early or late passage of 333-8-9 t cells. aav structural antigens were not detected in these cells unless the cells were coinfected with a helper adenovirus. in early passage 333-8-9 t cells were permissive to infections with ... | 1977 | 210924 |
| complementation of adeno-associated virus growth with temperature-sensitive mutants of human adenovirus types 12 and 5. | temperature-sensitive mutants of human adenovirus types 12 and 5, defective in viral dna synthesis, were able to support growth of adeno-associated virus type i at the non-permissive temperature. | 1975 | 213533 |
| [helper viruses of adeno-associated virus type 4 replication]. | in replication of adeno-associated virus type 4 (aav-4) the helper function may be performed by a non-defective virus from the same group of parvoviruses (kilham virus). the synthesis of aav-4 antigen was observed in a pig embryo kidney cell line, spev, chronically infected with kilham virus, strain rv-13, 45--52 passages. a one-day-old spev-kilham culture was infected with aav-4. the aav-4 antigen was detected by immunofluorescence at 6, 8, 12, 18 hours, 2, 3, 4, and 5 days after inoculation. d ... | 1979 | 217178 |
| parvovirus rna transcripts containing sequences not present in mature mrna: a method for isolation of putative mrna precursor sequences. | we report here a method of rna preparation that may enrich for precursor rna sequences and the results of an investigation of adeno-associated virus (aav) rna transcription that used this method. whole cells were lysed with detergent and high salt and separated into supernatant and pellet (crude chromatin) fractions. these fractions were then separately deproteinized by proteolytic digestion and phenol extractions. dna was removed from the preparation by two cycles of pancreatic dnase digestion ... | 1979 | 218213 |
| adeno-associated virus autointerference. | | 1979 | 218354 |
| adenovirus-associated virus polypeptides synthesized in cells coinfected with either adenovirus or herpesvirus. | | 1979 | 219599 |
| adeno-associated virus dna replication: nonunit-length molecules. | | 1979 | 219605 |
| defective-interfering particles of the human parvovirus adeno-associated virus. | | 1979 | 220782 |
| studies on the defectiveness of adeno-associated virus (aav). i. effects of phosphonoacetic acid and 2-deoxy-d-glucose on the replication of aav. | | 1979 | 222062 |
| studies on the defectiveness of adeno-associated virus (aav). ii. effect of growth in cv-1 cells on the replication of adeno-associated virus. | | 1979 | 222063 |
| mechanism of host restriction of adenovirus-associated virus replication in african green monkey kidney cells. | human adenovirus (ad) serotypes provide an early factor(s) that is necessary for adenovirus-associated virus (aav) multiplication in human cell lines. however, little, if any, aav production occurs in primary african green monkey kidney (agmk) cells co-infected with aav and a helper human ad (non-permissive infection), unless cells are additionally infected with sv40 (permissive infection). to determine the basis of the host restriction of aav replication in agmk cells, aav dna, rna and protein ... | 1979 | 225435 |
| isolation of an adeno associated virus from sheep. brief report. | an adenovirus and a spherical virus 20--24 nm diameter were isolated from ovine faeces. the small virus replicated in the nucleus, and was associated especially with the nucleolus. it haemagglutinated guinea pig and human erythrocytes, was thermostable and required an adenovirus for replication. it is concluded that this represents the first recorded isolate of an ovine aav. | 1979 | 226038 |
| [incidence of antibodies against adeno-associated viruses in commercial poultry]. | | 1979 | 226059 |
| adeno-associated virus dna replication. | | 1979 | 226321 |
| adeno-associated virus--an extreme state of viral defectiveness. | | 1979 | 228349 |
| inhibition and enhancement of avian adenovirus plaque production by heavy and light avian adenovirus-associated viral particles. | the effect of heavy and light avian adenovirus-associated viral (a-av) particles on the replication of several adenovirus serotypes was studied in chicken embryo kidney cells. there was a significant decrease (p less than 0.05) in the number and size of adenovirus-induced plaques at a-av multiplicities of infection greater than 40. enhancement of plaque production was observed when a-av multiplicities of infection were 1 to 40. there was a significant increase in the number and size of infective ... | 1979 | 229748 |
| adenovirus-associated virus structural protein sequence homology. | adenovirus-associated virus (aav) structural proteins (vp1, vp2, and vp3) have been examined to determine if areas of sequence homology exist between these three virion proteins. tryptic and chymotryptic maps have been produced which demonstrate extensive areas of sequence homology common to all three proteins. the amino acid compositions of the proteins were also determined and were found to be very similar. these data are consistent with the hypothesis that all three virion proteins arise eith ... | 1979 | 230301 |
| spliced adenovirus-associated virus rna. | we describe the structure of cytoplasmic rna species transcribed from the dna of adenovirus-associated virus, a defective parvovirus. the rna was hybridized with minus strand template dna and visualized in the electron microscope. alternatively, the dna.rna duplex molecules were digested with nuclease s1 or escherichia coli exonuclease vii and analyzed by agarose gel electrophoresis. a set of rna species was observed with 5' terminal at map positions 5, 13, 19, or 39 and a 3' terminus and poly(a ... | 1979 | 230481 |
| adeno-associated viruses. | | 1979 | 230727 |
| 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 |
| use of adeno-associated virus as a general transduction vector for mammalian cells. | | 1992 | 1316261 |
| in vitro replication of adeno-associated virus dna. | an in vitro assay for adeno-associated virus (aav) dna replication has been developed. the substrate is a plasmid containing the duplex form of aav dna in pbr322. the aav insert is excised or rescued from the plasmid by extracts of uninfected cells. replication was assayed by production of full-length excised aav dna resistant to dpn i digestion. the following results were obtained. (i) only extracts of cells coinfected with aav and adenovirus replicated the excised insert. (ii) density label ex ... | 1992 | 1316616 |
| analysis of mutations in adeno-associated virus rep protein in vivo and in vitro. | the adeno-associated virus (aav) rep protein is required for both viral dna replication and transactivation of the aav promoters. here we report the effects of mutations in the rep gene on transcription and replication in vivo and terminal repeat binding and terminal resolution site (trs) endonuclease activities in vitro. in all, we examined 10 in-frame deletions and 14 amino acid substitution mutations at eight positions. the point mutations were targeted to regions that are highly conserved am ... | 1992 | 1318396 |
| induction of differentiation-associated changes in established human cells by infection with adeno-associated virus type 2. | the nonpathogenic human defective parvovirus adeno-associated virus (aav) type 2 induced differentiation-associated antigens in cells of the human leukemia cell line hl60 (cd 67), as well as in two different lines of immortalized human keratinocytes, hacat and hpk ia cells (involucrin and cytokeratin 10). simultaneously, expression of the c-myc and c-myb oncogenes and the retinoblastoma gene was down regulated whereas c-fos expression increased in infected cells. these data point to the potentia ... | 1992 | 1318400 |
| inhibition of bovine papillomavirus plasmid dna replication by adeno-associated virus. | the helper-dependent human parvovirus adeno-associated virus type 2 (aav) inhibits both the oncogenic transforming abilities and the dna replication of its helper viruses, adenovirus (ad), and herpes simplex virus (hsv). as aav-2 also inhibits the transforming ability of bovine papillomavirus type 1 (bpv), aav-2 was assayed for its ability to inhibit bpv plasmid dna replication. here we find that the aav-2 rep78 gene is able to trans-inhibit bpv plasmid dna replication and that the aav-2 termina ... | 1992 | 1318608 |
| detection of adeno-associated virus type 2 in human peripheral blood cells. | the non-pathogenic human parvovirus, adeno-associated virus (aav) is helper virus-dependent. however, it integrates into the cellular genome in the absence of its helper viruses. therefore it could become a useful vector for gene therapy. previous studies and our own results have shown that 40 to 80% of adults are seropositive for aav and that seroconversion occurs during the first few years of life, but little is known about the route of natural infection with the virus. we used the polymerase ... | 1992 | 1321880 |
| regulated high level expression of a human gamma-globin gene introduced into erythroid cells by an adeno-associated virus vector. | gene therapy of severe hemoglobinopathies will require high-level expression of a transferred globin gene in erythroid cells. distant regulatory elements flanking the beta-globin gene cluster, the locus control region, are needed for appropriate expression. we have explored the use of a human parvovirus, the adeno-associated virus (aav), for globin gene transfer. the human a gamma-globin gene, linked to hypersensitivity site 2 from the locus control region of the beta-globin gene cluster, was su ... | 1992 | 1323131 |
| adeno-associated virus sensitizes hela cell tumors to gamma rays. | infection with the helper virus-dependent human parvovirus adeno-associated virus (aav) is known to interfere with cellular transformation in vitro and oncogenesis in vivo. here we report on sensitization to gamma irradiation by aav infection of cells in culture and of tumors established from hela cells grafted into immunodeficient (nude) mice: infection of hela cells with aav type 2 enhanced cell killing and reduced plating efficiency after irradiation compared with uninfected cells. similarly, ... | 1992 | 1323717 |
| gene expression from adeno-associated virus vectors in airway epithelial cells. | lung diseases such as cystic fibrosis (cf) might be treated by gene therapy using viral vectors delivered to the airway. one potential vector is the defective human parvovirus, adeno-associated virus (aav). we examined the aav p5 transcription promoter for gene expression in immortalized cell lines derived from the airway (ib3-1) or pancreas (cfpac-1) of cf patients. aav vectors expressing the prokaryotic genes cat (paavp5cat) or neo (paavp5neo) from the p5 promoter were evaluated after introduc ... | 1992 | 1325813 |
| modulation of the cellular phenotype by integrated adeno-associated virus. | the adeno-associated virus (aav) rep gene encodes a series of overlapping, multifunctional, nonstructural proteins (rep proteins) which regulate the viral life cycle and which are also capable of trans-regulating nonviral gene expressions (reviewed in berns, 1990, microbiol. rev. 54, 316-329). to investigate the expression of the aav rep gene in a cellular chromosomal context, sv40-transformed chinese hamster embryo (od4) cells were infected with an aav/neo hybrid virus and progeny resistant to ... | 1992 | 1326806 |
| cloning and integration of dna fragments in human cells via the inverted terminal repeats of the adeno-associated virus 2 genome. | in current systems for molecular cloning of eukaryotic genes, bacterial cells are routinely utilized as intermediate hosts. we investigated the possibility of using a viral system for cloning dna fragments independent of bacterial cell usage. in this report, we provide an alternative approach for molecular cloning of dna fragments in eukaryotic cells by utilizing the inverted terminal repeats (itrs) of the genome of a nonpathogenic human parvovirus, the adeno-associated virus 2 (aav). we constru ... | 1992 | 1327964 |
| adenovirus containing a deletion of the early region 2a gene allows growth of adeno-associated virus with decreased efficiency. | efficient growth of adeno-associated virus (aav) requires helper functions provided by a coinfecting adenovirus or herpesvirus. earlier studies using adenoviruses having temperature-sensitive lesions in the early region 2a gene (e2a) produced contradictory evidence regarding the role of the e2a 72-kda dna-binding protein (dbp) in allowing efficient aav growth. these disparate results may reflect varying levels of residual function in the temperature-sensitive dbp. we examined this issue using an ... | 1992 | 1329332 |
| 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 |
| dual-target inhibition of hiv-1 in vitro by means of an adeno-associated virus antisense vector. | an adeno-associated virus vector encoding an antisense rna was used to transduce stable intracellular resistance to human immunodeficiency virus-1 (hiv-1) in human hemopoietic and non-hemopoietic cell lines. the antisense targets are present in all hiv-1 transcripts and include the tar sequence, which is critical for transcription and virus replication, and the polyadenylation signal. cell lines expressing antisense rna showed up to 95 percent inhibition of gene expression directed by the hiv-1 ... | 1992 | 1359646 |
| adeno-associated virus vectors. | adeno-associated virus is a human parvovirus that integrates its dna genome into host cell chromosomes with very high efficiency. this suggests that adeno-associated virus may be a useful vector for human gene therapy. interest in adeno-associated virus vectors increased greatly in the last year following reports that adeno-associated virus genome integration may be site specific and occur at preferred sites in the human genome. several genes relevant to the treatment of genetic or infectious di ... | 1992 | 1369403 |
| 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 |
| the evolution of defective and autonomous parvoviruses. | because of the small size and genetic simplicity of small dna viruses, parvoviruses would appear to be excellent models for studying viral evolution and adaptation. in an earlier publication we hypothesized the evolution of sequences of cellular "junk" dna into protective interfering transposons. these transposons would interfere with invading pathogenic viruses by competing with the pathogen dna for replicative enzymes. we speculated that a small, defective parvovirus, the adeno-associated viru ... | 1991 | 1648152 |
| adeno-associated viruses having nonsense mutations in the capsid genes: growth in mammalian cells containing an inducible amber suppressor. | when an adeno-associated virus (aav) genome contained in a recombinant plasmid is transfected into adenovirus-infected cells, infectious aav particles are efficiently generated. we previously described the construction of a conditional lethal mutant of aav having an amber termination codon inserted in the rep gene. this mutant was propagated on a monkey kidney cell line (supd12) having an inducible amber suppressor trnaser. we now describe the construction and propagation of two additional condi ... | 1991 | 1651593 |
| construction and expression of a recombinant adeno-associated virus that harbors a human beta-globin-encoding cdna. | towards a goal of using recombinant adeno-associated viruses (aav) for the gene therapy of hemoglobinopathies we had previously constructed plasmid pav h beta g psi 1, which contained a human beta-globin-encoding cdna (hbb) downstream from the p40 promoter of aav2 dna [ohi et al., gene 89 (1990) 279-282]. transfection of the plasmid into human 293 cells (embryonal kidney cell line) resulted in the expression of hbb at the mrna level as well as rescue and replication of the recombinant aav genome ... | 1991 | 1655580 |
| biological and physicochemical characterization of the major (1.40) and minor (1.45) component of infectious avian adeno-associated virus. | two infectious components with buoyant densities of 1.40 g/cm3 and 1.45 g/cm3, designated as major (1.40) and minor (1.45) component, were detected by banding avian adeno-associated virus (aaav) isopycnically in cscl. in metrizamide, however, infectious aaav banded only as a single peak at a density of 1.32 g/cm3. biological as well as physicochemical properties of the two aaav components recovered from cscl density gradient were described. concerning the minor (1.45) component, three experiment ... | 1991 | 1656917 |
| replication of the human immunodeficiency virus type 1 is inhibited by the adeno-associated virus rep gene. | | 1991 | 1665443 |
| establishment of epithelial cell lines from human and mouse thymus immortalized by the 12s adenoviral e1a gene product. | to understand the role of thymic epithelial cells in the development of immature thymocytes, the establishment and cloning of thymic epithelial cell lines must be required. in the present study, we established human and mouse thymic epithelial cell lines through the immortalization by the transfection of cdna sequences of adenoviral e1a 12s mrna. this procedure resulted in the isolation of five stable cell lines (one human cell line and four mouse cell lines). these cell lines were positive in c ... | 1991 | 1723817 |
| 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 |
| a subset of herpes simplex virus replication genes provides helper functions for productive adeno-associated virus replication. | herpesviruses are helper viruses for productive adeno-associated virus (aav) replication. to analyze the herpes simplex virus type 1 (hsv-1) functions mediating helper activity, we coinfected hela cells with aav type 2 (aav-2) and different hsv-1 mutants defective in individual hsv replication genes. aav replication was fully accomplished in the absence of hsv dna replication and thus did not require expression of late hsv genes. in addition, hsv mutants lacking either the origin-binding protein ... | 1991 | 1850024 |
| infection with adeno-associated parvovirus leads to increased sensitivity of mammalian cells to stress. | infection of the sv40-transformed syrian hamster cells elona with adeno-associated parvovirus type 5 (aav-5) affects the cellular stress response. stress-induced growth delay becomes permanent and this promotes cell death. aav dna is replicated in the absence of helper virus. induction of an incomplete stress response by cycloheximide treatment still results in replication of viral dna but not in virus-mediated growth arrest. thus, the reactions that induce aav dna replication and that direct gr ... | 1991 | 1850906 |
| 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 |
| in vitro resolution of covalently joined aav chromosome ends. | we have developed an assay for a key step in the replication of adeno-associated virus (aav) dna. we demonstrate the covalently joined ends of linear aav dna can be resolved in vitro to the open duplex configuration. only extracts prepared from human cells that have been infected with both adenovirus and aav are capable of carrying out the reaction. the reaction is initiated by a site-specific and strand-specific endonucleolytic cut at a terminal resolution site near the end of the aav terminal ... | 1990 | 2153052 |
| site-specific integration by adeno-associated virus. | cellular sequences flanking integrated copies of the adeno-associated virus (aav) genome were isolated from a latently infected clonal human cell line and used to probe genomic blots derived from an additional 21 independently derived clones of human cells latently infected with aav. in genomic blots of uninfected human cell lines and of primary human tissue, each flanking-sequence probe hybridized to unique bands, but in 15 of the 22 latently infected clones the flanking sequences hybridized no ... | 1990 | 2156265 |
| mutation of a consensus purine nucleotide binding site in the adeno-associated virus rep gene generates a dominant negative phenotype for dna replication. | adeno-associated virus (aav) contains a multifunctional nonstructural gene, rep, which is required for aav dna replication and has pleiotropic effects on positive and negative regulation of gene expression. all of the parvovirus nonstructural genes contain a region of highly conserved amino acid homology. within this conserved region is the consensus sequence for a purine nucleotide binding site. we constructed a mutant aav having a mutation in this site by converting lysine 340 to histidine. th ... | 1990 | 2157057 |
| the adeno-associated virus rep gene suppresses herpes simplex virus-induced dna amplification. | herpes simplex virus (hsv) induces within the host cell genome dna amplification which can be suppressed by coinfection with adeno-associated virus (aav). to characterize the aav functions mediating this effect, cloned aav type 2 wild-type or mutant genomes were transfected into simian virus 40 (sv40)-transformed hamster cells together with the six hsv replication genes (encoding ul5, ul8, major dna-binding protein, dna polymerase, ul42, and ul52) which together are necessary and sufficient for ... | 1990 | 2159559 |
| inhibition of n-methyl-n'-nitro-n-nitrosoguanidine-induced methotrexate and adriamycin resistance in cho cells by adeno-associated virus type 2. | we studied the effects of helper-dependent parvovirus aav [adeno-associated virus] type 2 on carcinogen-inducible resistance to methotrexate (mtx) and adriamycin (adr) in chinese hamster ovary cells. both types of drug resistance were monitored by determination of the number of drug-resistant colonies normalized for the respective value of plating efficiency under non-selective conditions. treatment of cells with n-methyl-n'-nitro-n-nitrosoguanidine (mnng) drastically enhanced the frequency of r ... | 1990 | 2161803 |
| infection with adeno-associated virus type 5 inhibits mutagenicity of herpes simplex virus type 1 or 4-nitroquinoline-1-oxide. | infection with adeno-associated virus type 5 (aav-5) reduced the number of mutants arising in the hypoxanthine phosphoribosyltransferase locus of human rd 176 cells after infection with herpes simplex virus type 1 (hsv-1; partially inactivated) or 4-nitroquinoline-1-oxide (4-nqo). the mutation frequency was reduced by aav-5 infection from 11.4 to 1.8 after mutation with hsv-1 and from 3.2 to 2.5 when mutation was induced by 4-nqo. this was analyzed by determination of the number of cells resista ... | 1990 | 2166909 |
| 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 |
| 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 |
| 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 |
| targeted integration of adeno-associated virus (aav) into human chromosome 19. | a key feature in adeno-associated virus (aav) replication is efficient integration of the viral genome into host cell dna to establish latency when helper virus is absent. the steps involved in this process remain largely uncharacterized, even though aav integration was first documented 20 years ago. using a protein--dna binding method we isolated aav--cellular junction dna sequences. the cellular component hybridized to a single restriction fragment in the virus-free parental cell line, and als ... | 1991 | 1657596 |
| 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 |
| mapping and direct visualization of a region-specific viral dna integration site on chromosome 19q13-qter. | a human parvovirus, adeno-associated virus (aav), is unique among eukaryotic dna viruses in its ability to integrate site specifically into a defined region of human chromosome 19. in this study we used in situ hybridization to visualized directly the site of aav dna integration in latently infected human cell lines and normal human cells. | 1991 | 1653762 |
| adeno-associated virus rep proteins produced in insect and mammalian expression systems: wild-type and dominant-negative mutant proteins bind to the viral replication origin. | the adeno-associated virus (aav) rep gene proteins, rep78 and rep68, are required for replication of aav dna and bind to the aav replication origin. an aav genome having a lys340 to his (k340h) mutation in the consensus purine nucleotide binding site of the rep gene protein exhibited a dominant-negative phenotype for dna replication. we synthesized both wild-type and the k340h mutant rep78 protein in a baculovirus expression system. nuclear extracts of sf9 cells containing these proteins were ex ... | 1991 | 1651588 |
| 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 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 |
| status of ribozyme and antisense-based developmental approaches for anti-hiv-1 therapy. | | 1992 | 1340161 |
| characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus dna by non-homologous recombination. | the human parvovirus, adeno-associated virus (aav), has been shown to integrate preferentially into human chromosome 19 q13.3-qter. the human target sequence for aav integration (aavs1) was cloned and sequenced. by analysis of the proviral junctions it was determined that integration of the aav dna occurred via a non-homologous recombination pathway although there were either four or five identical nucleotides at the junctions. integration was a multistep, concerted process that resulted in cell ... | 1992 | 1334463 |
| 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 |
| 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 |
| complementation for replication by unrelated animal viruses containing dna genomes. | | 1987 | 2830477 |
| 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 |
| adeno-associated viruses inhibit sv40 dna amplification and replication of herpes simplex virus in sv40-transformed hamster cells. | coinfection with hsv-1 and any of the three defective parvoviruses aav-2, aav-3, or aav-5 reduces the hsv-1-induced amplification of sv40 dna in the sv40-transformed hamster cell lines co631 and elona. moreover, all three viruses significantly decrease hsv-1 replication. the effects are measurable to the same extent in both cell lines and are dependent on the quantity of aav dna molecules rather than on infectious aav particles. it seems unlikely that the inhibitions are due to simple competitio ... | 1988 | 2834875 |
| 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 |
| gene transfer in human lymphocytes using a vector based on adeno-associated virus. | adeno-associated virus is a nonpathogenic, dependent parvovirus that integrates at a specific site in human chromosome 19. we have used the inverted terminal repeats of the virus, which mediate integration, to establish a vector for gene transfer in human lymphocytes. a neomycin resistance gene has been stably introduced into nontransformed human t-cell clones and a subsequent analysis of the functional properties of the infected clone revealed no detectable alterations. rescue and replication o ... | 1992 | 1318074 |
| 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 |
| 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 |
| dissociation of carcinogen-induced sv40 dna-amplification and amplification of aav dna in a chinese hamster cell line. | time kinetics of aav-5 dna amplification induced by mnng in cells of a chinese hamster line (co631) and a syrian hamster line (elona) were compared to kinetics of sv40 dna amplification in these cells. aav-5 dna amplification starts in both lines about 8 hr following treatment of aav-infected cells with mnng. sv40 dna amplification is induced only in co631 cells. the time it becomes detectable varies but in no case was before 23 hr after mnng treatment. in co631 cells a second round of aav dna a ... | 1988 | 2842939 |
| expression and rescue of a nonselected marker from an integrated aav vector. | we used rep+ and rep- recombinant aav-plasmid vectors containing the nonselectable marker chloramphenicol acetyltransferase (cat) driven by the aav p40 promoter, and having a selectable marker, neo, inserted in the plasmid genome, and driven by a herpesvirus thymidine kinase gene promoter. each vector was transfected into human 293 cells or hela cells and the neo gene was used to select geneticin-resistant (genr) cells containing integrated vectors. the genr cells were then screened for expressi ... | 1988 | 2842941 |
| 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 |
| 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 |
| modification of some biological properties of hela cells containing adeno-associated virus dna integrated into chromosome 17. | parvoviruses are known to interfere with cellular transformation and carcinogenesis. since infecting adeno-associated virus (aav) frequently integrates its dna into the cellular genome, we analyzed whether this integration influences the transformed phenotype of the human tumor cell line hela. analysis of three independent hela cell clones with integrated aav dna (ha-3x, ha-16, and ha-28) revealed the following phenotypic changes of these cells: (i) reduced growth rate, (ii) increased serum requ ... | 1992 | 1313913 |
| 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 |
| immunologic and virologic status of multitransfused patients: role of type and origin of blood products. by the aids-hemophilia french study group. | an immunologic and virologic work-up was undertaken in 425 symptom-free multitransfused patients with hemophilias or hemoglobinopathies living in france. patients were entered into five groups according to the type of blood product they received: local factor viii, a mixture of local and imported factor viii, imported factor ix, local factor ix, washed red blood cells. the overall prevalence of igg antibodies to the lymphadenopathy-associated virus (lav) was 45%. the highest rate was observed in ... | 1985 | 2994780 |
| influence of adeno-associated virus on adherence and growth properties of normal cells. | it has been shown previously that infection of newly established cell cultures from malignant human tumors with adeno-associated parvovirus type 2 or type 5 results in growth arrest and cell death. here we report that the additionally observed antiproliferative effect on diploid human fibroblasts is transient and is connected to a reduced number of cells in s phase. progression through the cell cycle is disturbed either in g0/g1 or at the g1/s boundary, but an additional arrest in g2 cannot be e ... | 1992 | 1309911 |
| 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 |
| 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 |
| 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 |
| 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 |