RRC ID 34759
Author Nozaki S, Niki H, Ogawa T.
Title Replication initiator DnaA of Escherichia coli changes its assembly form on the replication origin during the cell cycle.
Journal J Bacteriol
Abstract DnaA is a replication initiator protein that is conserved among bacteria. It plays a central role in the initiation of DNA replication. In order to monitor its behavior in living Escherichia coli cells, a nonessential portion of the protein was replaced by a fluorescent protein. Such a strain grew normally, and flow cytometry data suggested that the chimeric protein has no substantial loss of the initiator activity. The initiator was distributed all over the nucleoid. Furthermore, a majority of the cells exhibited certain distinct foci that emitted bright fluorescence. These foci colocalized with the replication origin (oriC) region and were brightest during the period spanning the initiation event. In cells that had undergone the initiation, the foci were enriched in less intense ones. In addition, a significant portion of the oriC regions at this cell cycle stage had no colocalized DnaA-enhanced yellow fluorescent protein (EYFP) focus point. It was difficult to distinguish the initiator titration locus (datA) from the oriC region. However, involvement of datA in the initiation control was suggested from the observation that, in DeltadatA cells, DnaA-EYFP maximally colocalized with the oriC region earlier in the cell cycle than it did in wild-type cells and oriC concentration was increased.
Volume 191(15)
Pages 4807-14
Published 2009-8
DOI 10.1128/JB.00435-09
PII JB.00435-09
PMID 19502409
PMC PMC2715721
MeSH Bacterial Proteins / genetics Bacterial Proteins / metabolism Bacterial Proteins / physiology* Blotting, Western Cell Cycle / genetics Cell Cycle / physiology* DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism DNA-Binding Proteins / physiology* Escherichia coli / cytology Escherichia coli / genetics* Escherichia coli / metabolism* Luminescent Proteins / genetics Luminescent Proteins / metabolism Microscopy, Fluorescence Polymerase Chain Reaction Replication Origin / genetics Replication Origin / physiology*
IF 3.234
Times Cited 14
Prokaryotes E. coli ME9769 ME9770 ME9771 ME9772 ME9773 ME9774 ME9775 ME9776 ME9777 ME9778 ME9779