Infectious proviral DNA in human cells infected with transformation-defective type C viruses

Nicolson, Margery; Hariri, Fadi; Krempin, H.M.; McAllister, Robert M.; Gilden, R. V.

doi:10.1016/0042-6822(76)90273-7

Cited by 22 publications

(19 citation statements)

References 30 publications

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“…The present report also shows that the AKR cell DNA (9). However, Svoboda et al (24) reported that this treatment enhanced the transfection efficiency of the Rous sarcoma viral genome when DNA from XC cells was transfected onto chicken embryo fibroblasts.…”

Section: Akr Mulv or Rauscher Mulv Is Infectious With Or Withoutmentioning

confidence: 46%

“…The data are consistent with an IdUrd-sensitive restriction in the recipient NIH 3T3 cells that revents viral replication from endogenous AKR MuLV DNA Cut does not prevent viral replication from MuLV DNA of productively infected cells. Previous transfection studies have shown that retroviral DNA is infectious either as an unintegrated proviral DNA genome from acutely infected cells (1)(2)(3)(4) or as an integrated proviral DNA genome from chronically infected cells (5)(6)(7)(8)(9)(10). However, DNA from cells known to contain unexpressed but potentially infectious endogenous viral genomes is not infectious under the same transfection conditions (11).…”

Section: Akr Mulv or Rauscher Mulv Is Infectious With Or Withoutmentioning

confidence: 99%

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Infectious murine leukemia virus from DNA of virus-negative AKR mouse embryo cells.

Lowy¹

1978

Proc. Natl. Acad. Sci. U.S.A.

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DNA from virus-negative AKR mouse embryo cells is infectious for NIH 3T3 cells if the transfected cells are treated with 5-iododeoxyuridine (IdUrd) either before or after addition of the DNA. The virus isolated after transfection of the AKR DNA is an ecotropic murine leukemia virus (MuLV) Previous transfection studies have shown that retroviral DNA is infectious either as an unintegrated proviral DNA genome from acutely infected cells (1-4) or as an integrated proviral DNA genome from chronically infected cells (5-10). However, DNA from cells known to contain unexpressed but potentially infectious endogenous viral genomes is not infectious under the same transfection conditions (11). Thus, except for the demonstration that DNA from avian cells can complement the reverse transcriptase lesion in a temperature-sensitive mutant of Rous sarcoma virus (12), DNA from normal cells has not yet been shown to be infectious, despite genetic, biologic, and biochemical studies that indicate that endogenous retroviral genomes are present in cells from many species (13,14). The demonstration of complete retroviral expression from the DNA of normal cells would show directly that the cell DNA is potentially infectious. In addition, an infectivity assay for an endogenous viral genome would be useful in attempting to elucidate the cellular and viral mechanisms that restrict the expression of the viral genome in the endogenous state and permit its expression in the productively infected state.Although all inbred mice (Mus musculus) apparently contain endogenous retroviral genomes (15, 16) that are potentially infectious (17), strains vary markedly in their viral expression in vivo and in vitro (18)(19)(20)(21) Chemicals. IdUrd was purchased from Sigma and it was stored at 1 mg/ml in sterile phosphate-buffered saline at -20°C.DNA Isolation. Bulk cellular DNA was isolated as described (30) except that, prior to treatment with RNase, the ethanolprecipitated nucleic acid was wound on a rod instead of being concentrated by centrifugation. This modification was used because the centrifuged DNA was more difficult to redissolve than was the "spooled" DNA. This DNA had an average molecular weight of greater than 5 X 107. It was stored at 150-300 ,g/ml (20 A260 units/,ug DNA) at 4°C in 10 mM NaCl/10 mM Tris, pH 8.0 /0.5 mM EDTA. Stored DNA is still infectious after 1 year.Because it was critical that untreated AKR 2B cells not be producing virus at the time of DNA isolation, at least 2 X 106 AKR cells from the pool of cells used for each AKR DNA isolation were cocultivated with SC-1 cells, passaged at least twice, and found at that time to be negative for MuLV by the XC plaque test (31). IdUrd-treated AKR DNA was obtained after 24-hr treatment of AKR 2B cells with IdUrd at 40,g/ml. Incorporation of IdUrd into DNA was confirmed by CsCl density gradient centrifugation. Infectious center plating onto SC-1 cells Abbreviations: MuLV, murine leukemia virus; IdUrd, 5-iododeoxyuridine.

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Section: Akr Mulv or Rauscher Mulv Is Infectious With Or Withoutmentioning

confidence: 46%

Section: Akr Mulv or Rauscher Mulv Is Infectious With Or Withoutmentioning

confidence: 99%

Infectious murine leukemia virus from DNA of virus-negative AKR mouse embryo cells.

Lowy¹

1978

Proc. Natl. Acad. Sci. U.S.A.

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“…In order to obtain integrated DNA copies of these viral genomes, viruses ofthe three subtypes were used to infect dog or human cells. DNAs were extracted from the cells 6 weeks after infection and examined by restriction endonuclease blotting analysis with the U3 probe. Under these circumstances, only internal viral bands were visible because of the great heterogeneity ofjunction fragments after a mass infection.…”

Section: Resultsmentioning

confidence: 99%

“…DNAs were transfected into dog D17, feline embryo fibroblasts (AH 927), and human RD4 cells as described (5). Isolation of DNA from tissues and tissue culture cell lines was carried out as described (6).…”

mentioning

confidence: 99%

The U3 portion of feline leukemia virus DNA identifies horizontally acquired proviruses in leukemic cats.

Casey¹,

Roach²,

Mullins³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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The presence and location of DNA sequences related to the U3 and U5 portions ofthe infectious exogenous feline leukemia virus (FeLV) long terminal repeat (LTR) in various cat DNAs have been determined by hybridization experiments. In uninfected cat DNAs, the U5 LTR segment from the GardnerArnstein strain B virus is present at approximately 150 copies per cell. This level is approximately 10-fold greater than that of endogenous internal FeLV sequences. The U5 sequences differ in copy number and, to some extent, in location from one animal to another. For any one animal, the sequence organization ofthe U5

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“…In our initial experiments, however, RD-114 complementary DNA (cDNA) hybridized to DNA from the virus-infected human cell at a much slower rate -14-fold) than to normal cat cell DNA. Transfection of RD-114 using DNA from infected human cells has been achieved [5], whereas similar experiments with normal cat cells have yielded negative results [M.O. N icolson et al, unpublished data].…”

mentioning

confidence: 99%

Reiteration Frequency of Feline Type C Viral Genomes in Homologous and Heterologous Host Cell DNA

Okabe¹,

duBuy²,

Hatanaka³

et al. 1978

Intervirology

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Differences in the multiplicity of proviral sequences of two feline endogenous viruses, RD-114 and FeLV, in homologous and heterologous host DNA were examined by viral cDNA hybridization using cellular DNA fractionated with respect to reiteration frequency. The endogenous proviral DNAs in cat cells were fractionated predominantly with intermediate-repeated sequences, while RD-114 proviralDNA in virus-infected human cells was fractionated together with unique-sequence DNA. These differences were consistent with the results of kinetic analysis using unfractionated DNA.

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Infectious proviral DNA in human cells infected with transformation-defective type C viruses

Cited by 22 publications

References 30 publications

Infectious murine leukemia virus from DNA of virus-negative AKR mouse embryo cells.

Infectious murine leukemia virus from DNA of virus-negative AKR mouse embryo cells.

The U3 portion of feline leukemia virus DNA identifies horizontally acquired proviruses in leukemic cats.

Reiteration Frequency of Feline Type C Viral Genomes in Homologous and Heterologous Host Cell DNA

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