학위논문 (석사)-- 서울대학교 대학원 자연과학대학 생명과학부, 2017. 8. 황덕수.The centrosome serves as a major microtubule-organizing center (MTOC). The Cdc6 protein, which is a component of pre-replicative complex and a licensing factor for the initiation of chromosome replication, localizes to centrosomes during the S- and G2-phases of the cell cycle of human cells. We found that Cdc6 negatively regulates MTOC activity and the amounts of the integral proteins composing the pericentriolar material (PCM). Whereas Cdc6 depletion increased microtubule nucleation at the centrosomes, induction of Cdc6 in Cdc6-depleted cells reduced this increase. This increase and reduction correlated with the centrosomal intensities of PCM proteins, such as γ-tubulin, pericentrin, CDK5RAP2, and Cep192. These regulations required the ATPase activity as well as the centrosomal localization of Cdc6. These results suggest a novel Cdc6 function that regulates centrosome assembly and function.Introduction 1
Materials and Methods 3
DNA construction and transfection 3
Short interfering RNA (siRNA) transfection 3
Microtubule regrowth assay 4
Immunofluorescence microscopy 5
BrdU incorporation and fluorescence-activated cell sorting (FACS) analysis 5
Statistical analysis 6
Results 7
Cdc6 depletion increases microtubule regrowth originating from centrosomes 7
Centrosomal localization of Cdc6 protein reduces microtubule formation 8
ATPase activity of Cdc6 is necessary for the reduction of microtubule formation 18
Cdc6 reduces microtubule formation in S- and G2-phase centrosomes 27
Cdc6 controls microtubule nucleation 34
Cdc6 negatively regulates the levels of PCM proteins 37
Discussion 53
References 56
Abstract in Korean 68Maste
