Negative control of DNA replication by hydrolysis of ATP bound to DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli

被引：61

作者：

Mizushima, T ^{[1
]}

Nishida, S ^{[1
]}

Kurokawa, K ^{[1
]}

Katayama, T ^{[1
]}

Miki, T ^{[1
]}

Sekimizu, K ^{[1
]}

机构：

[1] KYUSHU UNIV,FAC PHARMACEUT SCI,FUKUOKA 81282,JAPAN

来源：

EMBO JOURNAL | 1997年 / 16卷 / 12期

关键词：

ATPase; DnaA; Escherichia coli; negative regulation; site-directed mutagenesis;

D O I：

10.1093/emboj/16.12.3724

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：

DnaA protein, the initiation factor for chromosomal DNA replication in Escherichia coli, is activated by ATP, ATP bound to DnaA protein is slowly hydrolyzed to ADP, but the physiological role of ATP hydrolysis is unclear We constructed, by site-directed mutagenesis, mutated DnaA protein with lower ATPase activity, and we examined its function in vitro and in vivo. The ATPase activity of purified mutated DnaA protein (Glu204-->Gln) decreased to one-third that of the wildtype DnaA protein. The mutation did not significantly affect the affinity of DnaA protein for ATP or ADP, The mutant dnaA gene showed lethality in wild-type cells but not in cells growing independently of the function of oriC. Induction of the mutated DnaA protein in wild-type cells caused an overinitiation of DNA replication. Our results lead to the thesis that the intrinsic ATPase activity of DnaA protein negatively regulates chromosomal DNA replication in E.coli cells.

引用

页码：3724 / 3730

页数：7

共 32 条

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