Identification of a preinitiation step in DNA replication that is independent of origin recognition complex and cdc6, but dependent on cdk2

被引:181
作者
Hua, XQH [1 ]
Newport, J [1 ]
机构
[1] Univ Calif San Diego, Dept Biol, La Jolla, CA 92093 USA
关键词
D O I
10.1083/jcb.140.2.271
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Before initiation of DNA replication, origin recognition complex (ORC) proteins, cdc6, and minichromosome maintenance (MCM) proteins bind to chromatin sequentially and form preinitiation complexes. Using Xenopus laevis egg extracts, we find that after the formation of these complexes and before initiation of DNA replication, cdc6 is rapidly removed from chromatin, possibly degraded by a cdk2-activated, ubiquitin-dependent proteolytic pathway. If this displacement is inhibited, DNA replication fails to initiate. We also find that after assembly of MCM proteins into preinitiation complexes, removal of the ORC from DNA does not block the subsequent initiation of replication. Importantly, under conditions in which both ORC and cdc6 protein are absent from preinitiation complexes, DNA replication is still dependent on cdk2 activity. Therefore, the final steps in the process leading to initiation of DNA replication during S phase of the cell cycle are independent of ORC and cdc6 proteins, but dependent on cdk2 activity.
引用
收藏
页码:271 / 281
页数:11
相关论文
共 57 条
[1]   ATP-DEPENDENT RECOGNITION OF EUKARYOTIC ORIGINS OF DNA-REPLICATION BY A MULTIPROTEIN COMPLEX [J].
BELL, SP ;
STILLMAN, B .
NATURE, 1992, 357 (6374) :128-134
[2]  
BROEK D, 1991, NATURE, V349, P388, DOI 10.1038/349388a0
[3]   DUAL FUNCTIONS OF CDC6 - A YEAST PROTEIN REQUIRED FOR DNA-REPLICATION ALSO INHIBITS NUCLEAR DIVISION [J].
BUENO, A ;
RUSSELL, P .
EMBO JOURNAL, 1992, 11 (06) :2167-2176
[4]   Role for a Xenopus Orc2-related protein in controlling DNA replication [J].
Carpenter, PB ;
Mueller, PR ;
Dunphy, WG .
NATURE, 1996, 379 (6563) :357-360
[5]   A MULTIUBIQUITIN CHAIN IS CONFINED TO SPECIFIC LYSINE IN A TARGETED SHORT-LIVED PROTEIN [J].
CHAU, V ;
TOBIAS, JW ;
BACHMAIR, A ;
MARRIOTT, D ;
ECKER, DJ ;
GONDA, DK ;
VARSHAVSKY, A .
SCIENCE, 1989, 243 (4898) :1576-1583
[6]   PURIFICATION OF AN MCM-CONTAINING COMPLEXES A COMPONENT OF THE DNA-REPLICATION LICENSING SYSTEM [J].
CHONG, JPJ ;
MAHBUBANI, HM ;
KHOO, CY ;
BLOW, JJ .
NATURE, 1995, 375 (6530) :418-421
[7]   An essential role for the Cdc6 protein in forming the pre-replicative complexes of budding yeast [J].
Cocker, JH ;
Piatti, S ;
Santocanale, C ;
Nasmyth, K ;
Diffley, JFX .
NATURE, 1996, 379 (6561) :180-182
[8]   The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts [J].
Coleman, TR ;
Carpenter, PB ;
Dunphy, WG .
CELL, 1996, 87 (01) :53-63
[9]   P25(RUM1) ORDERS S-PHASE AND MITOSIS BY ACTING AS AN INHIBITOR OF THE P34(CDC2) MITOTIC KINASE [J].
CORREABORDES, J ;
NURSE, P .
CELL, 1995, 83 (06) :1001-1009
[10]   Chromatin binding, nuclear localization and phosphorylation of Xenopus cdc21 are cell-cycle dependent and associated with the control of initiation of DNA replication [J].
Coue, M ;
Kearsey, SE ;
Mechali, M .
EMBO JOURNAL, 1996, 15 (05) :1085-1097