Baboon endogenous virus genome. I. Restriction enzyme map of the unintegrated DNA genome of a primate retrovirus

Unintegrated linear and circular forms of baboon endogenous type C virus M7 DNA were prepared from M7-infected cells by chromatography on hydroxyapatite columns, and the circular DNAs were purified in cesium chloride-ethidium bromide equilibrium density gradients. The circular DNAs were linearized by digestion with EcoRI, which had a unique site on the viral DNA. The linearized DNA was then inserted into AgtWES.XB at the EcoRI site and cloned in an approved EK2 host. Molecularly cloned full-length M7 DNA was restricted with BamHI, and the resulting five subgenomic fragments were then subcloned individually in plasmid pBR322. The organization and sites of integration of the approximately 100 copies of M7 DNA sequences endogenous to baboons were investigated by digesting the DNA with restriction enzymes and identifying the virus-specific fragments by hybridization to labeled probes made by using the molecularly cloned full-length and subgenomic fragments of the viral DNA. We found that most of the endogenous sequences had sizes and organizations similar to those of the unintegrated viral DNA and therefore approximately similar to the RNA of the infectious virus. A few of the multiple sequences had deletions in the 3' end (envelope region), and some of the sequences either lacked or contained modified BamHI restriction sites on the 5' end of the viral DNA. The endogenous viral DNA sequences were nontandem, uninterrupted, and colinear with the DNA of the infectious virus, and they were integrated at different sites in the baboon DNA, like the M7 proviral DNA sequences acquired upon infection.

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“…tion map of one of these recombinant clones is shown in Fig. 3; this map is similar to the M7 linear DNA map described recently (3,12,22).…”

Section: Resultssupporting

confidence: 70%

Organization of type C viral DNA sequences endogenous to baboons: analysis with cloned viral DNA

Battula¹,

Hager²,

Todaro³

1982

J Virol

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“…The following retroviruses were provided by the Viral Resources Laboratory, Frederick Cancer Research Center: BaEV (M7) from A204 cells; simian sarcoma virus/simian sarcoma-associated virus [SSV-1 (SSAV)] from NC-37 cells; gibbon ape leukemia virus (GaLV-H) from 6g-1 cells; SMRV from D-17 cells; MPMV from CMMT cells; RD-114 from RD cells; BALB-2 from Cf2Th cells; NZB from Cf2Th cells; endogenous virus of rhesus monkeys (MMC-1) from Cf2Th cells; R-MuLV from JLS-V9 cells; and feline leukemia virus (FeLV) (A, B, C) from RD cells (7). The procedure for isolation of 70S viral RNA was described previously (14). Randomly primed [32P]cDNA probes were synthesized from the 70S viral RNAs as reported earlier (14,38).…”

Section: Methodsmentioning

confidence: 99%

“…The procedure for isolation of 70S viral RNA was described previously (14). Randomly primed [32P]cDNA probes were synthesized from the 70S viral RNAs as reported earlier (14,38). Under these conditions, cDNA's prepared with BaEV or reticuloendotheliosis virus (REV-A) 70S RNA were judged to be representative probes since they hybridized to DNA fragments throughout their respective genomes.…”

Section: Methodsmentioning

confidence: 99%

“…BaEV clone DNAs were cleaved by standard single and double restriction enzyme digestions, electrophoresed, and blotted as previously described (14,34). From the known BaEV restriction map (13,14), the completeness of digestion was monitored simply by the appearance of the expected restriction fragments and the absence of incompletely digested ones. Unless otherwise stated, 0.1 1Lg of BaEV clone 7B2 DNA was loaded in each lane of the agarose gel.…”

Section: Methodsmentioning

confidence: 99%

“…We initially characterized the BaEV genome by restriction analysis of BaEV unintegrated linear DNA (14) and the related proviruses of baboon cells and tissues (13). The characterization was extended to other BaEV isolates by analysis of unintegrated BaEV DNAs (6) and through cloning of these DNAs into A vectors (27).…”

mentioning

confidence: 99%

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DNA Sequence Relationship of the Baboon Endogenous Virus Genome to the Genomes of Other Type C and Type D Retroviruses

Cohen¹,

Rice²,

Stephens³

et al. 1982

J Virol

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Baboon endogenous virus (BaEV) is a type C retrovirus present in multiple proviral copies in the DNA of baboons. Although interspecies antigenic determinants present on reverse transcriptase and gag proteins are shared among all mammalian type C viruses, no nucleic acid homology between BaEV and other type C viruses (except RD-114) has been found in conventional liquid hybridization experiments. In this study, we used restriction fragments of cloned BaEV DNA immobilized on nitrocellulose to test for relatedness with [ 32 P]cDNA's of various type C and type D viruses. We detected the following distant relationships previously found only through immunological and protein sequencing techniques: (i) eight type C viral cDNA's (the endogenous virus of rhesus monkeys, feline leukemia virus, simian sarcoma virus, gibbon ape leukemia virus, Rauscher murine leukemia virus, BALB-2, NZB, and RD-114) and two type D viral cDNA's (Mason-Pfizer monkey virus and squirrel monkey retrovirus) were able to hybridize with cloned BaEV DNA; (ii) the eight type C probes hybridized to restriction fragments spanning most of the BaEV genome, but only RD-114 hybridized to fragments within the 1.9 kilobases at the 3′ end of the genome; (iii) the two type D probes hybridized primarily to fragments within the 1.9 kilobases at the 3′ terminus and weakly or not at all elsewhere; and (iv) [ 32 P]cDNA's of several other oncornaviruses (mouse mammary tumor virus, equine infectious anemia virus, bovine leukemia virus, and reticuloendotheliosis virus) exhibited no homology with BaEV DNA. DNA sequence analysis has allowed us to orient the BaEV restriction map with the genetic map at both ends of the genome. Homologies between retroviral cDNA's and BaEV clone restriction fragments could thus be related to specific BaEV genes. Whereas type C cDNA's hybridized to fragments from gag, pol , and the pol-env junction, squirrel monkey retrovirus cDNA hybridized only to a fragment coding for the p15E portion of env. Mason-Pfizer monkey virus cDNA also hybridized within the p15E region, but exhibited homology to the 3′ half of gp70 as well. These results are discussed relative to previously reported antigenic relatedness of retroviral proteins. The data suggest that BaEV represents an important link in oncornavirus evolution.

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Visna virus DNA: Discovery of a novel gapped structure

et al. 1981

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Baboon endogenous virus genome. I. Restriction enzyme map of the unintegrated DNA genome of a primate retrovirus

Cited by 17 publications

References 25 publications

Organization of type C viral DNA sequences endogenous to baboons: analysis with cloned viral DNA

Organization of type C viral DNA sequences endogenous to baboons: analysis with cloned viral DNA

DNA Sequence Relationship of the Baboon Endogenous Virus Genome to the Genomes of Other Type C and Type D Retroviruses

Visna virus DNA: Discovery of a novel gapped structure

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