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PMID: 8349557 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genetic analysis of double-strand break repair in Escherichia coli.

Journal of bacteriology ·Vol. 175 ·No. 16 ·1993-08-00 ·Pages 5176-85

Takahashi NK, Kusano K, Yokochi T, Kitamura Y, Yoshikura H, Kobayashi I

Abstract

We had reported that a double-strand gap (ca. 300 bp long) in a duplex DNA is repaired through gene conversion copying a homologous duplex in a recB21 recC22 sbcA23 strain of Escherichia coli, as predicted on the basis of the double-strand break repair models. We have now examined various mutants for this repair capacity. (i) The recE159 mutation abolishes the reaction in the recB21C22 sbcA23 background. This result is consistent with the hypothesis that exonuclease VIII exposes a 3'-ended single strand from a double-strand break. (ii) Two recA alleles, including a complete deletion, fail to block the repair in this recBC sbcA background. (iii) Mutations in two more SOS-inducible genes, recN and recQ, do not decrease the repair. In addition, a lexA (Ind-) mutation, which blocks SOS induction, does not block the reaction. (iv) The recJ, recF, recO, and recR gene functions are nonessential in this background. (v) The RecBCD enzyme does not abolish the gap repair. We then examined genetic backgrounds other than recBC sbcA, in which the RecE pathway is not active. We failed to detect the double-strand gap repair in a rec+, a recA1, or a recB21 C22 strain, nor did we find the gap repair activity in a recD mutant or in a recB21 C22 sbcB15 sbcC201 mutant. We also failed to detect conservative repair of a simple double-strand break, which was made by restriction cleavage of an inserted linker oligonucleotide, in these backgrounds. We conclude that the RecBCD, RecBCD-, and RecF pathways cannot promote conservative double-strand break repair as the RecE and lambda Red pathways can.

MeSH Terms
Bacterial Proteins/genetics Crossing Over, Genetic DNA Damage DNA Repair/genetics DNA-Binding Proteins/genetics Escherichia coli/genetics Escherichia coli Proteins Exodeoxyribonuclease V Exodeoxyribonucleases/genetics Gene Conversion Mutation Rec A Recombinases/metabolism Recombination, Genetic SOS Response, Genetics
Chemicals
Bacterial Proteins DNA-Binding Proteins Escherichia coli Proteins RecO protein, E coli RecR protein, E coli recF protein, E coli RecR protein, Bacteria recF protein, Bacteria Rec A Recombinases Exodeoxyribonucleases RecJ protein, E coli recE protein, E coli recJ protein, Bacteria Exodeoxyribonuclease V exodeoxyribonuclease V, E coli
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Takahashi N K
Department of Bacteriology, Faculty of Medicine, University of Tokyo, Japan.
Kusano K
Yokochi T
Kitamura Y
Yoshikura H
Kobayashi I
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.16.5176-5185.1993
Published
1993-08-00
Pages
5176-85
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC204985
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.175.16.5176-5185.1993 PMC Full Text View journal details
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