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. 1979 Feb;76(2):967–971. doi: 10.1073/pnas.76.2.967

src gene product from different strains of avain sarcoma virus: Kinetics and possible mechanism of heat inactivation of protein kinase activity from cells infected by transformation-defective, temperature-sensitive mutant and wild-type virus

Helga Rübsamen 1, Robert R Friis 1, Heinz Bauer 1
PMCID: PMC383109  PMID: 218225

Abstract

Sera from certain rabbits bearing Schmidt-Ruppin strain Rous sarcoma virus (RSV)-induced tumors precipitated p60src from chicken cells transformed by the homologous virus as well as by other strains [Prague strain RSV, Bryan high-titer strain RSV, and Bratislava 77 strain of avain sarcoma virus (ASV)], the molecular weights (Mrs) ranging from 60,000 to 64,000. The p60src immunoprecipitated from cells transformed by each of these strains incorporated [γ-32P]ATP into the Mr 53,000 subunit of IgG, though with differing activities. No such protein kinase activity (ATP:protein phosphotransferase, EC 2.7.1.37) was observed when the following immunoprecipitates were used: from uninfected cells, from untransformed cells infected by Rous-associated virus, or from cells transformed by acute leukosis viruses, avian erythroblastosis virus, or myelocytoma virus 29. The kinase reaction had a pH optimum at pH 5.9 and an apparent Km for ATP of 4.9 ± 2 μM, and was dependent on Mg2+ (Kb = 46 ± 12 mM), for which Ca2+ was no substitute. The kinase was cyclic AMP independent.

In order to test whether the protein kinase reaction is directly catalyzed by p60src, we compared the in vitro temperature sensitivities of the kinase activities from cells infected by transformation-temperature-sensitive mutant and parental wild-type virus. The first-order rate constant for the inactivation of the kinase from extracts of cells infected by the mutant virus was 2-fold greater than that from cells infected by wild-type virus. This result implicates the protein kinase as an enzymatic activity of the src gene product, the p60src. Concomitant with the loss of the kinase activity by heat inactivation, p60src loses 60-70% of its phosphate content. The kinetics of dephosphorylation exactly parallel those for the inactivation of the kinase activity, suggesting that the p60src kinase is itself dependent on phosphorylation for its activity.

Keywords: src gene product p60src, immunological group specificity, phosphorylation of protein kinase

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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