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PMID: 11230142 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Physical interactions between DinI and RecA nucleoprotein filament for the regulation of SOS mutagenesis.

The EMBO journal ·Vol. 20 ·No. 5 ·2001-03-01 ·Pages 1192-202

Yasuda T, Morimatsu K, Kato R, Usukura J, Takahashi M, Ohmori H

Abstract

The Escherichia coli dinI gene is one of the LexA-regulated genes, which are induced upon DNA damage. Its overexpression conferred severe UV sensitivity on wild-type cells and resulted in the inhibition of LexA and UmuD processing, reactions that are normally dependent on activated RecA in a complex with single-stranded (ss)DNA. Here, we study the mechanism by which DinI inhibits the activities of RecA. While DinI neither binds to ssDNA nor prevents the formation of RecA nucleoprotein filament, it binds to active RecA filament, thereby inhibiting its coprotease activity but not the ATPase activity. Furthermore, even under in vitro conditions where UmuD cleavage dependent on RecA-ssDNA-adeno sine-5'-(3-thiotriphosphate) is blocked in the presence of DinI, LexA is cleaved normally. This result, taken together with electron microscopy observations and linear dichroism measurements, indicates that the ternary complex remains intact in the presence of DinI, and that the affinity to the RecA filament decreases in the order LexA, DinI and UmuD. DinI is thus suited to modulating UmuD processing so as to limit SOS mutagenesis.

MeSH Terms
Adenosine Triphosphate/analogs & derivatives,metabolism Bacterial Proteins/antagonists & inhibitors,metabolism Circular Dichroism DNA, Bacterial/metabolism,ultrastructure DNA, Single-Stranded/metabolism,ultrastructure DNA-Binding Proteins/antagonists & inhibitors,metabolism,ultrastructure DNA-Directed DNA Polymerase Enzyme Activation Enzyme Stability Escherichia coli/genetics,metabolism Escherichia coli Proteins Microscopy, Electron Mutagenesis/genetics Protein Binding Rec A Recombinases/antagonists & inhibitors,metabolism,ultrastructure SOS Response, Genetics/genetics Serine Endopeptidases/metabolism Succinimides/metabolism Temperature
Chemicals
Bacterial Proteins DNA, Bacterial DNA, Single-Stranded DNA-Binding Proteins Escherichia coli Proteins LexA protein, Bacteria Succinimides adenosine 5'-O-(3-thiotriphosphate) Adenosine Triphosphate Rec A Recombinases DNA-Directed DNA Polymerase UmuD protein, E coli Serine Endopeptidases dithiobis(succinimidylpropionate)
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Yasuda T
Institute for Virus Research, Kyoto University, Japan.
Morimatsu K
Kato R
Usukura J
Takahashi M
Ohmori H
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.5.1192
Published
2001-03-01
Pages
1192-202
Language
English
Region
England
NLM ID
8208664
PMCID
PMC145485
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.5.1192 PMC Full Text View journal details
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