Partial characterization of a Moloney murine sarcoma virus 85,000-dalton polypeptide whose expression correlates with the transformed phenotype in cells infected with a temperature-sensitive mutant virus

Horn, Jacqueline Peltier; Wood, Theodore; Blair, Donald G.; Arlinghaus, Ralph B.

doi:10.1016/0042-6822(80)90052-5

Cited by 34 publications

(46 citation statements)

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“…Anti-p30 recognizes P85gW"--"" whereas. anti-plO does not (7). In the presence of manganese chloride (lanes a and b), phosphorylation of Mr 85,000 protein was detected in the anti-p30 but not in the antiplO immunoprecipitates.…”

Section: Resultsmentioning

confidence: 98%

“…The Immune Complex Kinase Activity Correlates with the Presence of P859'9-' Previous studies have established that P855ag5'-is not detected at 390C, the temperature that is nonpermissive for fibroblast transformation (7,8). Therefore, if the Mr 85,000 protein phosphorylated in the immune complex kinase reaction is indeed P85gag-ms, it should be absent from immune complex kinase reaction products of tsllO-infected 6m2 cells maintained at 390C.…”

Section: Resultsmentioning

confidence: 99%

“…The expression of P85"ag"' in these cells is temperature dependent because only P589ag is detected in cells maintained at 39°C. Such cells grown at 39°C lose characteristics usually associated with the transformed state (7)(8)(9).…”

mentioning

confidence: 99%

“…The v-mos gene has an open reading frame allowing for the translation of a Mr42,000-44,000 protein which is expressed in trace amounts in cells transformed by the 124 strain of Mo-MuSV (5). Work in our laboratory has characterized a variant of Mo-MuSV-124, known as tsllO MuSV, that codes for a gag-mos polyprotein of Mr 85,000 termed P8599-"°as well as for a P58gag (6)(7)(8). A nonproducer cell clone transformed by tsllO MuSV expresses easily detectable levels of both proteins at 33°C.…”

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confidence: 99%

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P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

Kloetzer

Maxwell

Arlinghaus

1983

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

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A protein identified as P859'9-°was shown to be phosphorylated when immunoprecipitates from tsllO Moloney murine sarcoma virus transformed nonproducer cells (clone 6m2) were incubated with [y-32P]ATP. The in vitro-labeled 85,000-dalton phosphoprotein comigrated on NaDodSO4/polyacrylamide gels with authentic phosphorylated P859'9-°. Immunoprecipitates obtained with antisera prepared against Rauscher murine leukemia virus core protein p30 were active in the immune complex kinase assay but anti-murine leukemia virus plO precipitates were not. Previous studies have shown that anti-p30 but not antiplO antisera recognize P859'9-°. The 6m2 clone has been shown to express P85gag-"w8 at 330C but not at 39TC. Anti-p30 immune complexes from 6m2 cells maintained at 390C failed to phosphorylate the 85,000-dalton protein. Furthermore, the in vitro phosphorylated 85,000-dalton protein gave the same pattern of V8 protease-generated cleavage products as in vivo 32P-labeled P85gag-'.8, We conclude from these results that P85gag`m06 is phosphorylated in anti-p30 immune complex kinase reactions. Phosphoamino acid analyses indicated that the in vitro phosphorylated P85gag-tno contained phosphoserine and phosphothreonine. Our findings indicate that incubation of anti-p30 immunoprecipitates at 39°C drastically reduced, in a specific way, the kinase activity associated with P85gag-1l8. This result and other data suggest that the kinase is virus-encoded. Because P85gag-"1w8, but not Pr65gag, is phosphorylated in anti-p30 immunoprecipitates from MuLVMuSV tsllO producer cells, the kinase enzyme is associated with P85gag-' and not gag gene products. A second major polypeptide of the size of P58gag was also phosphorylated in anti-p30 immunoprecipitates from cells maintained at 33°C but not at 39°C. Since 6m2 cells at 39°C contain P58wag, this is also consistent with the kinase activity being associated with P85gag-11W.

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Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

Kloetzer

Maxwell

Arlinghaus

1983

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

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“…The expression of the transformation state has been extensively studied in the avian sarcoma virus system (11 390C. One such cell clone (6m2) was characterized; it does not produce virus at either temperature but contains viral-specified polyproteins (p) of Mr 58,000 (p58) and 85,000 (p85) (13,14). p85 is detected only at the transformed state and containsthe antigenic determinants of the viral core proteins, p15, p12, and p30, in addition to other unidentified peptide sequences.…”

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confidence: 99%

Sequence of events in the transformation process in cells infected with a temperature-sensitive transformation mutant of Moloney murine sarcoma virus.

Brown¹,

Horn²,

Wible³

et al. 1981

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

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Normal rat kidney cells infected with the temperature-sensitive transformation mutant ofMoloney murine sarcoma virus were used to study the biochemical and morphological changes that occur during transformation. The infected cells exhibited a normal morphology at the nonpermissive temperature (39C) and a transformed morphology at the permissive temperature (33°C Within 5 hr at the permissive temperature, the lacy network was still present but the microtubules were more diffusely stained and less discernible. By 48 hr, the microtubules were so diffuse that the lacy network could not be recognized. Alterations in the Factin cables did not occur until 24 hr after shifting the cells to the permissive temperature. Enucleation of the cells at the nonpermissive temperature and shifting the cytoplasts to the permissive temperature did not result in the synthesis of detectable p85 or in any alteration of the cytoplast morphology or microtubule complex, suggesting that the temperature-sensitive lesion affects some event occurring in the nucleus.

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Analysis of v‐mos encoded proteins in cells transformed by several related murine sarcoma viruses

Papkoff

Lai

Hunter

1982

J of Cellular Biochemistry

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We have used antisera against synthetic peptides to identify and characterize a 37,000 dalton v-mos encoded protein (p37mos) in cells transformed by M-MuSV 124. p37mos, a phosphoprotein, comprises only about 0.0005% of total cellular protein in cell lines transformed by M-MuSV 124. NIH 3T3 cells acutely infected with M-MuSV 124, however, contain 30-100-fold more p37mos. These elevated levels of p37mos correlate with striking morphological changes and cell death in the acutely infected cell population. Using the antipeptide antisera, we have extended the analysis of v-mos proteins to include several other MuSV variants that contain a similar v-mos gene to M-MuSV 124. With the exception of P85, the gag-mos fusion protein from ts110 MuSV, the v-mos gene of these variants is expressed as a 35,000-37,000 dalton protein (size depending on the particular virus).

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Partial characterization of a Moloney murine sarcoma virus 85,000-dalton polypeptide whose expression correlates with the transformed phenotype in cells infected with a temperature-sensitive mutant virus

Cited by 34 publications

References 29 publications

P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

Sequence of events in the transformation process in cells infected with a temperature-sensitive transformation mutant of Moloney murine sarcoma virus.

Analysis of v‐mos encoded proteins in cells transformed by several related murine sarcoma viruses

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Partial characterization of a Moloney murine sarcoma virus 85,000-dalton polypeptide whose expression correlates with the transformed phenotype in cells infected with a temperature-sensitive mutant virus

Cited by 34 publications

References 29 publications

P85 gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

P85 gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

Sequence of events in the transformation process in cells infected with a temperature-sensitive transformation mutant of Moloney murine sarcoma virus.

Analysis of v‐mos encoded proteins in cells transformed by several related murine sarcoma viruses

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P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity

P85 ^gag-mos encoded by ts110 Moloney murine sarcoma virus has an associated protein kinase activity