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EMBO reports 6, 8, 736–741 (2005)
doi:10.1038/sj.embor.7400467
Published online: 22 July 2005
Hda inactivation of DnaA is the predominant mechanism preventing hyperinitiation of Escherichia coli DNA replication
Johanna E Camara1*, Adam M Breier2*†, Therese Brendler3, Stuart Austin3, Nicholas R Cozzarelli2 & Elliott Crooke1
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1 Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington, District of Columbia 20007, USA
2 Molecular and Cell Biology, University of California, Berkeley, 16 Barker Hall, Berkeley, California 94720, USA
3 NCI-DBS, Frederick Cancer Research and Development Center, Box B, Building 539/223, Frederick, Maryland 21702, USA
To whom correspondence should be addressed
Elliott Crooke Tel: +1 202 687 1644; Fax: +1 202 687 7186; E-mail: crooke@georgetown.edu
† Present address: Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave 68-530, Cambridge, Massachusetts 02139, USA
* These authors contributed equally to this work
Received 5 January 2005; Accepted 2 June 2005; Published online 22 July 2005.
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Abstract
Initiation of DNA replication from the Escherichia coli chromosomal origin is highly regulated, assuring that replication occurs precisely once per cell cycle. Three mechanisms for regulation of replication initiation have been proposed: titration of free DnaA initiator protein by the datA locus, sequestration of newly replicated origins by SeqA protein and regulatory inactivation of DnaA (RIDA), in which active ATP-DnaA is converted to the inactive ADP-bound form. DNA microarray analyses showed that the level of initiation in rapidly growing cells that lack datA was indistinguishable from that in wild-type cells, and that the absence of SeqA protein caused only a modest increase in initiation, in agreement with flow-cytometry data. In contrast, cells lacking Hda overinitiated replication twofold, implicating RIDA as the predominant mechanism preventing extra initiation events in a cell cycle.
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