Document Detail


Multicopy plasmids are clustered and localized in Escherichia coli.
MedLine Citation:
PMID:  11274369     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We localized the multicopy plasmid RK2 in Escherichia coli and found that the number of fluorescent foci observed in each cell was substantially less than the copy number of the plasmid, suggesting that many copies of RK2 are grouped into a few multiplasmid clusters. In minimal glucose media, the majority of cells had one or two foci, with a single focus localized near midcell, and two foci near the 1/4 and 3/4 cell positions. The number of foci per cell increased with cell length and with growth rate, and decreased upon entering stationary phase, suggesting a coordination of RK2 replication or segregation with the bacterial cell cycle. Time-lapse microscopy demonstrated that partitioning of RK2 foci is achieved by the splitting of a single focus into two or three smaller foci, which are capable of separating with rapid kinetics. A derivative of the high-copy-number plasmid pUC19 containing the lacO array was also localized by tagging with GFP-LacI. Whereas many of the cells contained numerous, randomly diffusing foci, most cells exhibited one or two plasmid clusters located at midcell or the cell quarter positions. Our results suggest a model in which multicopy plasmids are not always randomly diffusing throughout the cell as previously thought, but can be replicated and partitioned in clusters targeted to specific locations.
Authors:
J Pogliano; T Q Ho; Z Zhong; D R Helinski
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2001-03-27
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  98     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2001 Apr 
Date Detail:
Created Date:  2001-04-11     Completed Date:  2001-05-10     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4486-91     Citation Subset:  IM    
Affiliation:
Division of Biology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92130-0322, USA.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics
Base Sequence
DNA Primers
Escherichia coli / genetics*
Escherichia coli Proteins*
Green Fluorescent Proteins
Lac Repressors
Luminescent Proteins / genetics
Plasmids*
Repressor Proteins / genetics
Subcellular Fractions / metabolism
Grant Support
ID/Acronym/Agency:
AI-07194/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA Primers; 0/Escherichia coli Proteins; 0/Lac Repressors; 0/LacI protein, E coli; 0/Luminescent Proteins; 0/Repressor Proteins; 147336-22-9/Green Fluorescent Proteins
Comments/Corrections

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