The β-sliding clamp directs the localization of HdaA to the replisome in Caulobacter crescentus.

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Serval ID
serval:BIB_3FDD7A393EDE
Type
Article: article from journal or magazin.
Collection
Publications
Title
The β-sliding clamp directs the localization of HdaA to the replisome in Caulobacter crescentus.
Journal
Microbiology
Author(s)
Fernandez-Fernandez C., Grosse K., Sourjik V., Collier J.
ISSN
1465-2080 (Electronic)
ISSN-L
1350-0872
Publication state
Published
Issued date
2013
Peer-reviewed
Oui
Volume
159
Number
Pt 11
Pages
2237-2248
Language
english
Abstract
The initiation of chromosome replication is tightly regulated in bacteria to ensure that it takes place only once per cell cycle. In many proteobacteria, this process requires the ATP-bound form of the DnaA protein. The regulatory inactivation of DnaA (RIDA) facilitates the conversion of DnaA-ATP into replication-inactive DnaA-ADP, thereby preventing overinitiation. Homologues of the HdaA protein, together with the β-clamp of the DNA polymerase (DnaN), are required for this process. Here, we used fluorescence resonance energy transfer experiments to demonstrate that HdaA interacts with DnaN in live Caulobacter crescentus cells. We show that a QFKLPL motif in the N-terminal region of HdaA is required for this interaction and that this motif is also needed to recruit HdaA to the subcellular location occupied by the replisome during DNA replication. An HdaA mutant protein that cannot colocalize or interact with DnaN can also not support the essential function of HdaA. These results suggest that the recruitment of HdaA to the replisome is needed during RIDA in C. crescentus, probably as a means to sense whether chromosome replication has initiated before DnaA becomes inactivated. In addition, we show that a conserved R145 residue located in the AAA+ domain of HdaA is also needed for the function of HdaA, although it does not affect the interaction of HdaA with DnaN in vivo. The AAA+ domain of HdaA may therefore be required during RIDA after the initial recruitment of HdaA to the replisome by DnaN.
Pubmed
Web of science
Open Access
Yes
Create date
23/08/2013 11:07
Last modification date
20/08/2019 14:37
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