[Analysis of transient G1/S arrest in E1A+E1B-19kDa transformed cells after ionizing radiation].
Expression of human adenovirus type 5 E1A oncogene in normal rodent cells leads to disruption of the G1/S cell cycle arrest realization in response to DNA damage. It has been shown here that rat embryo fibroblasts transformed by E1Aad5 oncogene in complementation with E1B-19 kDa gene realize the irradiation-induced transient G1/S arrest, which depends on selective suppression of CyclinE-Cdk2 activity despite functional inactivation of p21Waf1 inhibitor. Inhibitor p21Waf1 is not revealed in complexes with cyclins E and A in E1A + E1B-19 kDa transformants, however, it is not due to p21Waf1 interaction with E1A oncoproteins, because the E1A-p21Waf1 complex formation in E1A + cHa-ras transformants does not prevent the high level of CycIE, A-p21Waf1 association. In the case of p21Waf1 inactivation, the main way of cyclin-kinase activity regulation in E1A + E1B-19 kDa cells may be Cdk2 phosphorylation. However, irradiation of E1A + E1B-19 kDa transformed cells induces no changes in CAK (Cdk7-associated) kinase activity and in the protein level of Cdc25A phosphatase, which are responsible for activating Thr160 phosphoralation and Tyr15 dephosphorylation on Cdk2. Using phospho-Tyr15-Cdk2 specific antibodies, no increase of phosphorylation at Tyr15 position on immunoprecipitated Cdk2 was detected after irradiation. It seems likely that in the case of inactivated inhibitor p21Waf1 the transient G1/S block after irradiation in E1A + E1B-19 kDa transformants depends on suppression of Cycl-E-Cdk2 activity caused by inhibition of Thr160 Cdk2 phosphorylation, but his occurs with the involvement of other kinases rather than CAK.
Brichkina
A I
AI
Institute of Cytology RAS, St. Petersburg. aibri@mail.ru
Aksenov
N D
ND
Pospelov
V A
VA
Pospelova
T V
TV
D017500:Adenovirus E1A Proteins / Q000096:biosynthesis / Q000235:genetics; D017501:Adenovirus E1B Proteins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D042846:CDC2-CDC28 Kinases / Q000032:analysis; D002461:Cell Line, Transformed / Q000166:cytology / Q000528:radiation effects*; D002471:Cell Transformation, Neoplastic / Q000528:radiation effects*; D019925:Cyclin A / Q000032:analysis; D019926:Cyclin B / Q000032:analysis; D051357:Cyclin-Dependent Kinase 2; D004249:DNA Damage; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000166:cytology / Q000528:radiation effects; D016193:G1 Phase; D010766:Phosphorylation; D051381:Rats; D016196:S Phase
D017500:Adenovirus E1A Proteins; D017501:Adenovirus E1B Proteins; D019925:Cyclin A; D019926:Cyclin B; D042846:CDC2-CDC28 Kinases; C495892:Cdk2 protein, rat; D051357:Cyclin-Dependent Kinase 2