The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA

Masayuki Su'etsugu, Yuji Harada, Kenji Keyamura, Chika Matsunaga, Kazutoshi Kasho, Yoshito Abe, Tadashi Ueda, Tsutomu Katayama

研究成果: ジャーナルへの寄稿記事

13 引用 (Scopus)

抄録

DnaA activity for replication initiation of the Escherichia coli chromosome is negatively regulated by feedback from the DNA-loaded form of the replicase clamp. In this process, called RIDA (regulatory inactivation of DnaA), ATP-bound DnaA transiently assembles into a complex consisting of Hda and the DNA-clamp, which promotes inter-AAA+ domain association between Hda and DnaA and stimulates hydrolysis of DnaA-bound ATP, producing inactive ADP-DnaA. Using a truncated DnaA mutant, we previously demonstrated that the DnaA N-terminal domain is involved in RIDA. However, the precise role of the N-terminal domain in RIDA has remained largely unclear. Here, we used an in vitro reconstituted system to demonstrate that the Asn-44 residue in the N-terminal domain of DnaA is crucial for RIDA but not for replication initiation. Moreover, an assay termed PDAX (pull-down after cross-linking) revealed an unstable interaction between a DnaA-N44A mutant and Hda. In vivo, this mutant exhibited an increase in the cellular level of ATP-bound DnaA. These results establish a model in which interaction between DnaA Asn-44 and Hda stabilizes the association between the AAA+ domains of DnaA and Hda to facilitate DnaA-ATP hydrolysis during RIDA.

元の言語英語
ページ(範囲)3183-3195
ページ数13
ジャーナルEnvironmental Microbiology
15
発行部数12
DOI
出版物ステータス出版済み - 12 1 2013

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inactivation
Adenosine Triphosphate
hydrolysis
DNA
Hydrolysis
mutants
chromosome
assay
Adenosine Diphosphate
crosslinking
Chromosomes
Escherichia coli
mutant
chromosomes
assays

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Ecology, Evolution, Behavior and Systematics

これを引用

The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA. / Su'etsugu, Masayuki; Harada, Yuji; Keyamura, Kenji; Matsunaga, Chika; Kasho, Kazutoshi; Abe, Yoshito; Ueda, Tadashi; Katayama, Tsutomu.

:: Environmental Microbiology, 巻 15, 番号 12, 01.12.2013, p. 3183-3195.

研究成果: ジャーナルへの寄稿記事

Su'etsugu, Masayuki ; Harada, Yuji ; Keyamura, Kenji ; Matsunaga, Chika ; Kasho, Kazutoshi ; Abe, Yoshito ; Ueda, Tadashi ; Katayama, Tsutomu. / The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA. :: Environmental Microbiology. 2013 ; 巻 15, 番号 12. pp. 3183-3195.
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AU - Ueda, Tadashi

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