Murine leukemia virus (T-8)-transformed cells: identification of a precursor polyprotein containing gag gene-coded proteins (p15 and p12) and a nonstructural component

Sacks, Thomas L.; Reynolds, Fred H.; Deobagkar, Dileep N.; Stephenson, John

doi:10.1128/jvi.27.3.809-814.1978

Cited by 27 publications

(14 citation statements)

References 25 publications

(28 reference statements)

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“…Recently, mink cells were transformed into non-producers by Abelson leukemia virus (Reynolds et al, 1978) and T-8 virus (Staal et a/., 1977) [a strain of mink cell focus-inducing virus (MCF) derived from the thymuses of preleukemic and leukemic AKR mice]. They appear to be genetic recombinants between different endogenous murine type-C viruses (Elder et af., 1977).…”

Section: Discussionmentioning

confidence: 99%

Moloney lymphoma cells express a polyprotein contanining the GAG gene‐coded p15 and the moloney leukemia virus‐induced cell surface antigen (MCSA)

Karande

Yefenof

Fenyõ

et al. 1979

Intl Journal of Cancer

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The relationship between MCSA and the viral structural proteins in YAC Moloney lymphoma cells was further investigated by using membrane immunofluorescence and immunoadsorbent columns. As previously observed, MCSA showed only minimal capping and thereby differed in behaviour from gp70, p30 and p12 virion antigens. Whereas antibody-induced capping of gp70 did not change the membrane distribution of MCSA, co-capping was observed between MCSA and p30 and p12 gag protein antigens. This indicated that, whereas MCSA is distinct from gp70, it is linked to p30 and p12 on the membrane of living cells. It was then attempted to isolate MCSA on anti-p30 and anti-p15 immunoadsorbent columns from solubilized YAC cells. Contrary to what was expected, no MCSA was found to the anti-p30 column. On the other hand, a part (10-15%) of the MCSA was bound to the anti-p15 column, indicating that MCSA is linked to the gag p15.

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Section: Discussionmentioning

confidence: 99%

Moloney lymphoma cells express a polyprotein contanining the GAG gene‐coded p15 and the moloney leukemia virus‐induced cell surface antigen (MCSA)

Karande

Yefenof

Fenyõ

et al. 1979

Intl Journal of Cancer

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“…In the present study, we have further characterized the 110,000 Mr polyprotein encoded by the T-8 isolate of MuLV (21). This highly phosphorylated viral-encoded protein is shown to contain amino terminal gag gene structural components but to lack immunologic cross-reactivity with other known AKR-MuLV translational products.…”

Section: Discussionmentioning

confidence: 88%

“…This is supported by the ability of such isolates, designated as mink cell focus forming (MCF) viruses, to induce morphologic alteration of mink cells in tissue culture (29). The more recent isolation of transforming variants from thymus cell-derived MCF virus stocks, one of which has been shown to be replication-defective (20) and to encode, as its major translation product, a 1 I0,000 Mr polyprotein (21), suggested a model analogous to that described for other mammalian RNA transforming viruses such as avian acute leukemia virus (4,7,10), several independent isolates of FeSV (5,6), and AbLV (8,9).…”

Section: Discussionmentioning

confidence: 99%

“…Cell lines were maintained in Dulbecco's modification of Eagle's medium (DME) supplemented with 10% calf serum (Colorado Serum Co., Denver, Co.). Cells used included a fetal mink lung cell line, CCL64, obtained from the American Culture Collection, Rockville, Md., and subclones of CCL64 nonproductively transformed by either T-8 virus (21) or AbLV (9) designated 64T8 and 64Ab-2, respectively. A Gardner FeSV transformed subclone of CCL64, designated 64F3, was generously provided by G. J. Todaro (National Cancer Institute, Bethesda, Md.).…”

Section: Methodsmentioning

confidence: 99%

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Identification of tryptic peptides unique to a 110,000-molecular weight polyprotein encoded by the T-8 isolate of murine leukemia virus.

Ven¹,

Reynolds²,

Blomberg³

et al. 1980

The Journal of Experimental Medicine

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Mink cells nonproductively-infected with the weakly-transforming T-8 isolate of murine leukemia virus (MuLV) express a 110,000 mol wt polyprotein designated T-8 P110. By immunoprecipitation analysis, T-8 P110 is shown to contain AKR-MuLV amino terminal gag gene-specific components (p15, p12) but to lack p30, p10, gp70, and p15(E) antigenic determinants. These observations are further substantiated by tryptic peptide analysis indicating T-8 P110 to share approximately six lysine-containing tryptic peptides with AKR-MuLV Pr65gag, and none with AKr-MuLV Pr82env. Furthermore, of seven methionine-containing T-8 P110 tryptic peptides, at least four can be conclusively shown not to be present in either AKr-MuLV Pr180gag/pol or Pr82env. A clonal mink cell line nonproductively infected by T-8, and expressing high levels of P110, although not morphologically transformed, is shown to lack elevated levels of tyrosine-specific protein kinase activity and reduction of epidermal growth factor binding sites characteristic of cells transformed by many other RNA-transforming viruses. These findings argue either that the T-8 viral genome contains acquired cellular sequences encoding a portion of P110, or that T-8 P110 represents an inphase deletion of AKR-MuLV Pr180gag/pol with extensive posttranlational modification and that an as yet unidentified protein is responsible for T-8 associated transformation.

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“…An increasing number of mammalian type C transforming viruses in addition to FeSV have been shown to code for 110,000 to 130,000 Mr polyproteins with both structural and nonstructural components (11,13,14,22). Such virusencoded polyproteins are of interest in view of the possible involvement of their nonstructural components in transformation.…”

Section: J Virolmentioning

confidence: 99%

Feline sarcoma virus-coded polyprotein: enzymatic cleavage by a type C virus-coded structural protein

Khan¹,

Stephenson²

1979

J Virol

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A purified 15,000-molecular-weight (Mr) Prague strain Rous sarcoma virus gag gene-coded structural protein, p15, was shown to enzymatically cleave the previously described 130,000 Mr feline sarcoma virus-coded polyprotein, Prl30. Cleavage products included proteins ranging in molecular weight from 12,000 to 110,000. The specificity of this cleavage reactivity was indicated by the fact that, under similar conditions, neither purified type C viral structural proteins nor nonviral proteins such as bovine serum albumin were cleaved to significant extents. Moreover, feline leukemia virus PrW5gag was efficiently cleaved, resulting in the generation of proteins of 30,000 (p30), 15,000 (p15), 12,000 (p12), and 10,000 (plO) Mr. Using enzymatically (p15) treated feline sarcoma virus Prl30 as starting material, we were able to purify a major 72,000 Mr cleavage product and to show it to contain the previously described feline sarcoma virus-coded nonstructural component.

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Murine leukemia virus (T-8)-transformed cells: identification of a precursor polyprotein containing gag gene-coded proteins (p15 and p12) and a nonstructural component

Cited by 27 publications

References 25 publications

Moloney lymphoma cells express a polyprotein contanining the GAG gene‐coded p15 and the moloney leukemia virus‐induced cell surface antigen (MCSA)

Moloney lymphoma cells express a polyprotein contanining the GAG gene‐coded p15 and the moloney leukemia virus‐induced cell surface antigen (MCSA)

Identification of tryptic peptides unique to a 110,000-molecular weight polyprotein encoded by the T-8 isolate of murine leukemia virus.

Feline sarcoma virus-coded polyprotein: enzymatic cleavage by a type C virus-coded structural protein

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