Document Detail


Replication initiator DnaA of Escherichia coli changes its assembly form on the replication origin during the cell cycle.
MedLine Citation:
PMID:  19502409     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Shingo Nozaki; Hironori Niki; Tohru Ogawa
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-06-05
Journal Detail:
Title:  Journal of bacteriology     Volume:  191     ISSN:  1098-5530     ISO Abbreviation:  J. Bacteriol.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-07-14     Completed Date:  2009-08-05     Revised Date:  2013-06-02    
Medline Journal Info:
Nlm Unique ID:  2985120R     Medline TA:  J Bacteriol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4807-14     Citation Subset:  IM    
Affiliation:
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya, Japan.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics,  metabolism,  physiology*
Blotting, Western
Cell Cycle / genetics,  physiology*
DNA-Binding Proteins / genetics,  metabolism,  physiology*
Escherichia coli / cytology,  genetics*,  metabolism*
Luminescent Proteins / genetics,  metabolism
Microscopy, Fluorescence
Polymerase Chain Reaction
Replication Origin / genetics,  physiology*
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA-Binding Proteins; 0/DnaA protein, Bacteria; 0/Luminescent Proteins; 0/yellow fluorescent protein, Bacteria
Comments/Corrections

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