Nearest neighbour analysis of MCM protein complexes in Drosophila melanogaster

被引:32
作者
Crevel, G
Ivetic, A
Ohno, K
Yamaguchi, M
Cotterill, S
机构
[1] Univ London St Georges Hosp, Sch Med, Dept Biochem, London SW17 0RE, England
[2] Aichi Canc Ctr, Res Inst, Div Biochem, Chikusa Ku, Nagoya, Aichi 4648681, Japan
关键词
D O I
10.1093/nar/29.23.4834
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The MCM proteins are a group of six proteins whose action is vital for DNA replication in eukaryotes. It has been suggested that they constitute the replicative helicase, with a subset of the proteins forming the catalytic helicase (MCM4,6,7) while the others have a loading or control function. In this paper we show that all six MCM proteins are present in equivalent amounts in soluble extracts and on chromatin. We have also analysed soluble and chromatin-associated MCM protein complexes under different conditions. This suggests that all six MCM proteins are always found in a complex with each other, although the interaction between the individual MCM proteins is not equivalent as stringent salt conditions are able to break the intact complex into a number of stable subcomplexes. These data contribute to the ongoing debate about the nature of MCM complexes, supporting the hypothesis that they act as a heterohexamer rather than as a number of different subcomplexes. Finally, using protein-protein cross-linking we have shown that MCM2 interacts directly with MCM5 and MCM6; MCM5 with MCM3 and MCM2; and MCM6 with MCM2 and MCM4. This provides the first direct information about specific subunit contacts in the MCM complex.
引用
收藏
页码:4834 / 4842
页数:9
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