title: Phosphorylation of cyclin O, a novel cyclin family protein containing a cyclin-like domain, is involved in the activation of cyclin-dependent kinase 2.
abstract: Cell cycles, ordered series of events modulating cell growth and division, are tightly regulated by complexes containing cyclin-dependent kinases (CDKs) and cyclins. Cyclin O is a novel cyclin family protein which interacts with CDK2. However, the molecular effects of cyclin O on the activity of CDK2 have not been fully evaluated. In this study, an interaction between cyclin O and CDK2 was identified by co-immunoprecipitation and the effect of cyclin O on the kinase activity of CDK2 was investigated using cyclin O point mutants. Co-immunoprecipitation was achieved using using HEK293 human embryonic kidney cells which were transiently transfected with vectors expressing cyclin O and CDK2, which revealed that cyclin O interacted with CDK2, particularly with the active form of endogenous CDK2. Cyclin O was expressed as several different bands with molecular weights between 45 and 50 kDa, possibly due to different post-translational modifications. When co-expressed with CDK2, cyclin O appeared as a band with a molecular weight of 50 kDa. Treatment with calf intestinal phosphatase reduced the intensity of the uppermost band. Mass spectroscopic analysis of cyclin O co-expressed with CDK2 revealed that the 81st serine residue of cyclin O was phosphorylated. Thein vitrokinase activity of CDK2 phosphorylating histone H1 was markedly increased in the cells overexpressing cyclin O. This activity was reduced in cells overexpressing cyclin O, in which the 81st serine had been replaced with alanine (S81A). These results suggest that cyclin O is a novel cyclin family protein that regulates CDK2 kinase activity, which is mediated by the phosphorylation of the 81st serine residue of cyclin O.
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
authors.forename: DO Hyung
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
authors.forename: Jong-Hwa
authors.initials: JH
authors.lastname: Park
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
authors.forename: Bora
authors.initials: B
authors.lastname: Lee
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
authors.forename: Kyoung Ok
authors.initials: KO
authors.lastname: Jang
authors.affiliation: Department of Genetic Engineering and Graduate School of Biotechnology, Kyung Hee University, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Korea.
authors.forename: In Sik
authors.initials: IS
authors.lastname: Chung
authors.affiliation: Department of Advanced Technology Fusion, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Ye Sun
authors.initials: YS
authors.lastname: Han
keywords: cyclin O; cyclin-dependent kinase 2; kinase activity; phosphorylation