Document Detail

Measuring chromosome dynamics on different time scales using resolvases with varying half-lives.
MedLine Citation:
PMID:  15853889     Owner:  NLM     Status:  MEDLINE    
The bacterial chromosome is organized into multiple independent domains, each capable of constraining the plectonemic negative supercoil energy introduced by DNA gyrase. Different experimental approaches have estimated the number of domains to be between 40 and 150. The site-specific resolution systems of closely related transposons Tn3 and gammadelta are valuable tools for measuring supercoil diffusion and analysing bacterial chromosome dynamics in vivo. Once made, the wild-type resolvase persists in cells for time periods greater than the cell doubling time. To examine chromosome dynamics over shorter time frames that are more closely tuned to processes like inducible transcription, we constructed a set of resolvases with cellular half-lives ranging from less than 5 min to 30 min. Analysing chromosomes on different time scales shows domain structure to be dynamic. Rather than the 150 domains detected with the Tn3 resolvase, wild-type cells measured over a 10 min time span have more than 400 domains per genome equivalent, and some gyrase mutants exceed 1000.
Richard A Stein; Shuang Deng; N Patrick Higgins
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular microbiology     Volume:  56     ISSN:  0950-382X     ISO Abbreviation:  Mol. Microbiol.     Publication Date:  2005 May 
Date Detail:
Created Date:  2005-04-27     Completed Date:  2005-09-20     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  8712028     Medline TA:  Mol Microbiol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1049-61     Citation Subset:  IM    
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MeSH Terms
Chromosomes, Bacterial / metabolism*
DNA Gyrase / genetics,  metabolism
Enzyme Stability
Nucleic Acid Conformation*
Recombinases / genetics,  metabolism*
Recombination, Genetic
Salmonella typhimurium / enzymology,  genetics
Time Factors
Grant Support
Reg. No./Substance:
0/Recombinases; EC Gyrase

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