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PMID: 10197989 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The RecBC enzyme loads RecA protein onto ssDNA asymmetrically and independently of chi, resulting in constitutive recombination activation.

Genes & development ·Vol. 13 ·No. 7 ·1999-04-01 ·Pages 901-11

Churchill JJ, Anderson DG, Kowalczykowski SC

Abstract

Double-strand DNA break repair and homologous recombination in Escherichia coli proceed by the RecBCD pathway, which is regulated by cis-acting elements known as chi sites. A crucial feature of this regulation is the RecBCD enzyme-directed loading of RecA protein specifically onto the 3'-terminal, chi-containing DNA strand. Here we show that RecBC enzyme (lacking the RecD subunit) loads RecA protein constitutively onto the 3'-terminal DNA strand, with no requirement for chi. This strand is preferentially utilized in homologous pairing reactions. We propose that RecA protein loading is a latent property of the RecBCD holoenzyme, which is normally blocked by the RecD subunit and is revealed following interaction with chi.

MeSH Terms
Blotting, Southern DNA Repair DNA, Single-Stranded/metabolism Escherichia coli/genetics Escherichia coli Proteins Exodeoxyribonuclease V Exodeoxyribonucleases/metabolism Genes, Bacterial Models, Genetic Oligonucleotide Probes Rec A Recombinases/metabolism Recombination, Genetic Time Factors
Chemicals
DNA, Single-Stranded Escherichia coli Proteins Oligonucleotide Probes Rec A Recombinases Exodeoxyribonucleases exodeoxyribonuclease I Exodeoxyribonuclease V exodeoxyribonuclease V, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Churchill J J
Biochemistry and Molecular Biology Graduate Group, University of California, Davis, California 95616-8665 USA.
Anderson D G
Kowalczykowski S C
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.13.7.901
Published
1999-04-01
Pages
901-11
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316600
Subset
IM
Grants
NIGMS NIH HHS · R01 GM041347 · United States
NIGMS NIH HHS · T32 GM007377 · United States
NIGMS NIH HHS · GM-41347 · United States
NIGMS NIH HHS · T32-GM-07377 · United States
External Links
PubMed source DOI: 10.1101/gad.13.7.901 PMC Full Text View journal details
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