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J. Bacteriol. doi:10.1128/JB.01294-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Dynamic association of the replication initiator and transcription factor DnaA with the Bacillus subtilis chromosome during replication stress

Adam M. Breier and Alan D. Grossman*

Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139

* To whom correspondence should be addressed. Email: adg{at}mit.edu.


   Abstract

DnaA functions as both a transcription factor and the replication initiator in bacteria. We characterized the DNA binding dynamics of DnaA on a genomic level. Based on crosslinking and immunoprecipitation (ChIP) data, DnaA binds at least 17 loci, 15 of which are regulated transcriptionally in response to inhibition of replication (replication stress). Six loci, each of which has a cluster of at least 9 potential DnaA binding sites, had significant increases in binding by DnaA when replication was inhibited, indicating that the association of DnaA with at least some of its target sites is altered after replication stress. When replication resumed from oriC after inhibition of replication initiation, these high levels of binding decreased rapidly at origin-proximal and origin-distal regions, well before a replication fork could pass through each of the regulated regions. These findings indicate that there is rapid signaling to decrease activation of DnaA during replication and that interaction between DnaA bound at each site and the replication machinery is not required for regulation of DnaA activity in response to replication stress.







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