Study of pp60 v-src protein kinase activity in synchronized chicken embryo fibroblasts infected with Rous Sarcoma Virus

Abstract

Chicken embryo fibroblast (CEF) cultures, synchronized by the addition of serum to stationary cells, were exposed to Schmidt-Ruppin strain of Rous Sarcoma Virus (SR-RSV) and the appearance of pp60 v-src protein kinase activity was examined through the cell cycle. In cells infected either at the beginning or at the end of G1, the onset of pp60 v-src protein kinase activity was coincidental, closely following mitosis, with a delay between the infection of cells with SR-RSV and the appearance of protein kinase activity of about 20 and 16 h, respectively. In cells infected during the S phase this delay was 16 h, as observed for late G1 cells. These experiments show that the activity of pp60 v-src protein kinase, which cannot be detected before the first mitosis following infection does not depend on G1. The aphidicolin prevented protein kinase activity if added before or at the beginning of S phase, but not if added later, which is presumably related to the inhibition of S phase, required for provirus integration. The use of colcemid, which suppresses cell division, did not inhibit but delayed the appearance of protein kinase activity. These results show that the synthesis of an active oncogene product, such as pp60 v-src protein kinase, depends on both S phase and mitosis.