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PMID: 15531636 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

RecA-dependent mutants in Escherichia coli reveal strategies to avoid chromosomal fragmentation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 46 ·2004-11-16 ·Pages 16262-7

Kouzminova EA, Rotman E, Macomber L, Zhang J, Kuzminov A

Abstract

RecA- and RecBC-catalyzed repair in eubacteria assembles chromosomes fragmented by double-strand breaks. We propose that recA mutants, being unable to repair fragmented chromosomes, depend on various strategies designed to avoid chromosomal fragmentation. To identify chromosomal fragmentation-avoidance strategies, we screened for Escherichia coli mutants synthetically inhibited in combination with recA inactivation by identifying clones unable to lose a plasmid carrying the recA(+) gene. Using this screen, we have isolated several RecA-dependent mutants and assigned them to three distinct areas of metabolism. The tdk and rdgB mutants affect synthesis of DNA precursors. The fur, ubiE, and ubiH mutants are likely to have increased levels of reactive oxygen species. The seqA, topA mutants and an insertion in smtA perturbing the downstream mukFEB genes affect nucleoid administration. All isolated mutants show varying degree of SOS induction, indicating elevated levels of chromosomal lesions. As predicted, mutants in rdgB, seqA, smtA, topA, and fur show increased levels of chromosomal fragmentation in recBC mutant conditions. Future characterization of these RecA-dependent mutants will define mechanisms of chromosomal fragmentation avoidance.

MeSH Terms
Adenosine Triphosphatases/genetics Bacterial Outer Membrane Proteins/genetics Chromosomes, Bacterial/genetics DNA Fragmentation DNA Helicases/genetics DNA Repair/genetics DNA-Binding Proteins/genetics Escherichia coli/genetics,metabolism Escherichia coli Proteins/genetics Genes, Bacterial Mutation Plasmids/genetics SOS Response, Genetics
Chemicals
Bacterial Outer Membrane Proteins DNA-Binding Proteins Escherichia coli Proteins SeqA protein, E coli Adenosine Triphosphatases DNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kouzminova Elena A
Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, IL 61801-3709, USA. kuzminov@life.uiuc.edu
Rotman Ella
Macomber Lee
Zhang Jian
Kuzminov Andrei
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0405943101
Email
kuzminov@life.uiuc.edu
Published
2004-11-16
Epub
2004-00-05
Pages
16262-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC528955
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0405943101 PMC Full Text View journal details
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