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PMID: 19451222 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Reconstitution of initial steps of dsDNA break repair by the RecF pathway of E. coli.

Genes & development ·Vol. 23 ·No. 10 ·2009-05-15 ·Pages 1234-45

Handa N, Morimatsu K, Lovett ST, Kowalczykowski SC

Abstract

The RecF pathway of Escherichia coli is important for recombinational repair of DNA breaks and gaps. Here ;we reconstitute in vitro a seven-protein reaction that recapitulates early steps of dsDNA break repair using purified RecA, RecF, RecO, RecR, RecQ, RecJ, and SSB proteins, components of the RecF system. Their combined action results in processing of linear dsDNA and its homologous pairing with supercoiled DNA. RecA, RecO, RecR, and RecJ are essential for joint molecule formation, whereas SSB and RecF are stimulatory. This reconstituted system reveals an unexpected essential function for RecJ exonuclease: the capability to resect duplex DNA. RecQ helicase stimulates this processing, but also disrupts joint molecules. RecO and RecR have two indispensable functions: They mediate exchange of RecA for SSB to form the RecA nucleoprotein filament, and act with RecF to load RecA onto the SSB-ssDNA complex at processed ssDNA-dsDNA junctions. The RecF pathway has many parallels with recombinational repair in eukaryotes.

MeSH Terms
DNA Breaks, Double-Stranded DNA Repair/genetics DNA-Binding Proteins/metabolism Escherichia coli/genetics,metabolism Escherichia coli Proteins/metabolism RecQ Helicases/metabolism
Chemicals
DNA-Binding Proteins Escherichia coli Proteins SSB protein, E coli recF protein, E coli RecQ protein, E coli RecQ Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Handa Naofumi
Department of Microbiology, University of California at Davis, Davis, Calfironia 95616, USA.
Morimatsu Katsumi
Lovett Susan T
Kowalczykowski Stephen C
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
1549-5477
DOI
10.1101/gad.1780709
Published
2009-05-15
Pages
1234-45
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC2685532
Subset
IM
Grants
NIGMS NIH HHS · R01 GM051753 · United States
NIGMS NIH HHS · GM-51753 · United States
NIGMS NIH HHS · R01 GM062653 · United States
NIGMS NIH HHS · R01 GM043889 · United States
NIGMS NIH HHS · GM-62653 · United States
NIGMS NIH HHS · GM-43889 · United States
NIGMS NIH HHS · R37 GM062653 · United States
External Links
PubMed source DOI: 10.1101/gad.1780709 PMC Full Text View journal details
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