Document Detail

DNA nicks inflicted by restriction endonucleases are repaired by a RecA- and RecB-dependent pathway in Escherichia coli.
MedLine Citation:
PMID:  10510229     Owner:  NLM     Status:  MEDLINE    
Two mutants of the EcoRI endonuclease (R200K and E144C) predominantly nick only one strand of the DNA substrate. Temperature sensitivity of the mutant enzymes allowed us to study the consequences of inflicting DNA nicks at EcoRI sites in vivo. Expression of the EcoRI endonuclease mutants in the absence of the EcoRI methyltransferase induces the SOS DNA repair response and greatly reduces viability of recA56, recB21 and lexA3 mutant strains of Escherichia coli. In parallel studies, overexpression of the EcoRV endonuclease in cells also expressing the EcoRV methyltransferase was used to introduce nicks at non-cognate EcoRV sites in the bacterial genome. EcoRV overproduction was lethal in recA56 and recB21 mutant strains and moderately toxic in a lexA3 mutant strain. The toxic effect of EcoRV overproduction could be partially alleviated by introduction into the cells of multiple copies of the E. coli DNA ligase gene. These observations suggest that an increased number of DNA nicks can overwhelm the repair capacity of DNA ligase, resulting in the conversion of a proportion of DNA nicks into DNA lesions that require recombination for repair.
J Heitman; T Ivanenko; A Kiss
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular microbiology     Volume:  33     ISSN:  0950-382X     ISO Abbreviation:  Mol. Microbiol.     Publication Date:  1999 Sep 
Date Detail:
Created Date:  1999-11-04     Completed Date:  1999-11-04     Revised Date:  2008-08-29    
Medline Journal Info:
Nlm Unique ID:  8712028     Medline TA:  Mol Microbiol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1141-51     Citation Subset:  IM    
Departments of Genetics, Pharmacology and Cancer Biology, Microbiology, and Medicine and the Howard Hughes Medical Institute, Duke University Medical Center, Box 3546, 322 Carl Building, Durham, NC 27710, USA.
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MeSH Terms
DNA Damage
DNA Ligases / genetics,  metabolism
DNA Repair / genetics,  physiology*
DNA, Bacterial / genetics,  metabolism*
Deoxyribonuclease EcoRI / genetics,  metabolism
Deoxyribonucleases, Type II Site-Specific / metabolism
Escherichia coli / genetics,  metabolism*
Escherichia coli Proteins*
Exodeoxyribonuclease V
Exodeoxyribonucleases / genetics,  metabolism
Genes, Bacterial
Rec A Recombinases / genetics,  metabolism
SOS Response (Genetics)
Site-Specific DNA-Methyltransferase (Adenine-Specific) / metabolism
Grant Support
Reg. No./Substance:
0/DNA, Bacterial; 0/Escherichia coli Proteins; EC 2.1.1.-/DNA modification methylase EcoRV; EC DNA-Methyltransferase (Adenine-Specific); EC 2.7.7.-/Rec A Recombinases; EC 3.1.-/Exodeoxyribonucleases; EC V; EC V, E coli; EC 3.1.21.-/Deoxyribonuclease EcoRI; EC, Type II Site-Specific; EC type II deoxyribonucleases; EC 6.5.1.-/DNA Ligases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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