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PMID: 22581769 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Topological characterization of the DnaA-oriC complex using single-molecule nanomanipuation.

Nucleic acids research ·Vol. 40 ·No. 15 ·2012-08-00 ·Pages 7375-83

Zorman S, Seitz H, Sclavi B, Strick TR

Abstract

In most bacteria, the timing and synchrony of initiation of chromosomal replication are determined by the binding of the AAA(+) protein DnaA to a set of high- and low-affinity sites found within the origin of chromosomal replication (oriC). Despite the large amount of information on the role and regulation of DnaA, the actual structure of the DnaA-oriC complex and the mechanism by which it primes the origin for the initiation of replication remain unclear. In this study, we have performed magnetic tweezers experiments to investigate the structural properties of the DnaA-oriC complex. We show that the DnaA-ATP-oriC complex adopts a right-handed helical conformation involving a variable amount of DNA and protein whose features fit qualitatively as well as quantitatively with an existing model based on the crystal structure of a truncated DnaA tetramer obtained in the absence of DNA. We also investigate the topological effect of oriC's DNA unwinding element.

MeSH Terms
Adenosine Triphosphate/metabolism Bacterial Proteins/chemistry,metabolism Biomechanical Phenomena DNA, Bacterial/chemistry DNA-Binding Proteins/chemistry,metabolism Escherichia coli/genetics Mutation Origin Recognition Complex/chemistry,genetics Replication Origin
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins DnaA protein, Bacteria OriC chromosomal replication origin Origin Recognition Complex Adenosine Triphosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zorman Sylvain
Institut Jacques Monod, CN RS UMR 7592, Univ Paris Diderot, Sorbonne Paris Cité, F-75205 Paris, France.
Seitz H
Sclavi B
Strick T R
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gks371
Published
2012-08-00
Epub
2012-00-11
Pages
7375-83
Language
English
Region
England
NLM ID
0411011
PMCID
PMC3424547
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gks371 PMC Full Text View journal details
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