Adhesion Plaques of Rous Sarcoma Virus-Transformed Cells Contain the src Gene Product (original) (raw)
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Comparison of the expression of the src gene of Rous sarcoma virus in vitro and in vivo
Journal of Virology
We have compared the polypeptide products of the src gene of several strains of Rous sarcoma virus produced by in vitro translation of heat-denatured 70S virion RNA in the nuclease-treated reticulocyte lysate with those present in chick cells transformed by these viruses. We have done this by immunoprecipitation, using sera from rabbits injected at birth with Schmidt-Ruppin Rous sarcoma virus. In vitro translation results in the synthesis of at least nine polypeptides which appear to be encoded by the src gene. These range in size from 17,000 to 60,000 daltons. The sera from tumor-bearing rabbits precipitated these polypeptides arising from the in vitro translation of RNA from Schmidt-Ruppin Rous sarcoma virus of both subgroup A and subgroup D and from one stock of Prague Rous sarcoma virus of subgroup C. In each case, all of this family of related polypeptides could be precipitated except the smallest, the 17,000-dalton polypeptide. No precipitation of analogous polypeptides resulting from the translation of RNA from other strains of Rous sarcoma virus was observed. Cells transformed by these three strains of Rous sarcoma virus contain easily detectable amounts of a polypeptide, p608rc essentially identical to the 60,000-dalton in vitro product. With one exception, they do not contain significant amounts of polypeptides analogous to the smaller in vitro products which can be precipitated by these sera. Cells transformed by one stock of Schmidt-Ruppin Rous sarcoma virus of subgroup A did contain a 39,000-dalton polypeptide, which was related, by peptide mapping, to the 60,000-dalton polypeptide and was similar in size to a precipitable in vitro product. The 60,000-dalton polypeptide present in transformed cells appeared to be phosphorylated 10 to 25 min after its synthesis, metabolically very stable, and not derived from a precursor polypeptide. All immunoprecipitates from transformed cells which contained p6Osrc also contained an 80,000-dalton phosphoprotein. This polypeptide is unrelated to p6Osrc, as determined by peptide mapping, and may well be a host cell polypeptide which is specifically associated with p608rc.
In vitro translation yields a possible Rous sarcoma virus src gene product
Proceedings of the National Academy of Sciences, 1977
In vitro translation of Rous sarcoma virus (RSV) virion RNA in the messenger-dependent reticulocyte lysate system yielded polypeptides that were not synthesized by translation of RNA from a transformation-defective deletion mutant of RSV. These RSV-specific products migrated on sodium dodecyl sulfate/polyacrylamide gels as two doublets of approximately 25,000 and 17,000 daltons. Synthesis of these proteins was not sensitive to inhibition by m7GTP; however, synthesis of the 76,000-dalton precursor of the internal structural proteins was sensitive to inhibition by m7GTP. Tryptic peptide maps showed the 25,000-and 17,000-dalton proteins to be related to one another but to be distinct from the 76,000-dalton protein.
Characterization of Rous sarcoma virus src gene products synthesized in vitro. J Virol
Journal of Virology
The cell-free synthesis of three major proteins from virion RNA of nondefective Rous sarcoma virus (RSV), but not from RNA of transformation-defective deletion mutants, has been observed. The apparent molecular weights of these transformation-specific proteins are approximately 60,000 (60K), 25K, and 17K. Tryptic maps of methionine-containing peptides revealed the 17K, 25K, and 60K proteins to be overlapping in sequence. However, only partial homology was observed Ibetween the 17K, 25K, and 60K proteins synthesized from Schmidt-Ruppin strain, subgroup D, RSV RNA and those synthesized from Prague strain, subgroup B, RSV RNA. About half of the methionine peptides in the Schmidt-Ruppin strain, subgroup D, 60K protein were shared with the Prague strain, subgroup D, 60K protein, and the rest were distinct to each. The virion RNAs coding for the 60K, 25K, and 17K proteins were found to be polyadenylated and to sediment with maximal mRNA activity at about 23, 19 to 20, and 18S, respectively. In addition, transformation-specific proteins with molecular weights of 39K and 33K were observed by in vitro synthesis. These proteins are also related to the 60K, 25K, and 17K proteins and were synthesized from polyaden-
Product of in vitro translation of the Rous sarcoma virus src gene has protein kinase activity
Journal of Virology, 1979
In vitro translation of Rous sarcoma virus virion RNA resulted in the synthesis of a protein kinase which, when immunoprecipitated with antitumor serum, phosphorylated the immunoglobulin heavy chain. Even though in vitro translation of virion RNA resulted in the synthesis of a number of polypeptides which were recognized by antitumor serum, control experiments demonstrated that an immunoprecipitable protein kinase activity was found only when an immunoprecipitable p60src, the polypeptide product of the src gene, was synthesized. A protein kinase with similar properties was therefore intimately associated with p60src which was synthesized in vitro in the reticulocyte lysate, just as it is with p60src which is obtained from transformed chick and mammalian cells. It is therefore highly unlikely that this association is artifactual. ts NY68 is a mutant of Rous sarcoma virus which is able to transform cells at 36 but not at 41 degrees C. In vitro translation of ts NY68 virion RNA at 30 d...
Characterization of Rous sarcoma virus src gene products synthesized in vitro
Journal of Virology, 1978
The cell-free synthesis of three major proteins from virion RNA of nondefective Rous sarcoma virus (RSV), but not from RNA of transformation-defective deletion mutants, has been observed. The apparent molecular weights of these transformation-specific proteins are approximately 60,000 (60K), 25K, and 17K. Tryptic maps of methionine-containing peptides revealed the 17K, 25K, and 60K proteins to be overlapping in sequence. However, only partial homology was observed between the 17K, 25K and 60K proteins synthesized from Schmidt-Ruppin strain, subgroup D, RSV RNA and those synthesized from Prague strain, subgroup B, RSV, RNA. About half of the methionine peptides in the Schmidt-Ruppin strain, subgroup D, 60K protein were shared with the Prague strain, subgroup D, 60K protein, and the rest were distinct to each. The virion RNAs coding for the 60K, 25K, and 17K proteins were found to be polyadenylated and to sediment with maximal mRNA activity at about 23, 19 to 20, and 18S, respectively...
Immunohistochemical detection of the gene product of Rous Sarcoma virus in human brain tumors
Brain Research, 1985
We have studied 108 cases of brain tumors by immunohistochemical staining utilizing an antiserum against the src gene product and GFA protein in order to elucidate the relationship between the brain tumors and the transforming genes. While the normal human brain tissues were not positive with the antiserum against the src gene product, several astrocytomas were clearly positive and other brain tumors were negative with the same. Src-gene product related peptide was detected in some of the human astrocytomas. 0006-8993/85/$03.30 (~) 1985 Elsevier Science Publishers B.V. (Biomedical Division)
Proceedings of the National Academy of Sciences, 1980
All vertebrate cells have been shown to contain a gene, sarc, that has some homology with the transforming gene of Rous sarcoma virus, src. We have com ared the polypeptide products of the sarc gene, p608arc, of human, mouse, and chicken cells with the polymorphic polypeptide product of the src gene, p60'rc, of several strains of Rous sarcoma virus by two-dimensional peptide mapping. p60sarc from chicken cells was clearly related to every viral p608'7. Eleven of its 13 methionine-containing tryptic peptides were present in some viral p60Orc. Conversely, the other two peptides were not present in any p60'tc we have examined so far. The 11 peptides from p60'7"c of chickens that were shared with viral p6OOrc, however, were not all present in any single viral p6087'c. These 11 peptides most closely resemble those in the p60'rcs of B77 virus and the Prague strain of Rous sarcoma virus. These data
Proceedings of the National Academy of Sciences, 1990
A retroviral promoter is sufficient to convert proto-src to a transforming gene that is distinct from the src gene of Rous sarcoma virus (proto-src recombinant virus/mutation enhances transforming function/recombinant cancer gene hypothesis) ABSTRACT The src genes of four natural isolates of avian sarcoma viruses differ from cellular proto-src in two genetic substitutions: the promoter of the cellular gene is replaced by a retroviral counterpart, and at least six codons from the 3' terminus: are replaced by retroviral or heterologous cellderived elements. Since virus constructs with a complete protosrc coding region failed to transform avian cells but acquired transforming function by point mutations of various codons, it