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PMID: 1438275 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Activity of the purified mutagenesis proteins UmuC, UmuD', and RecA in replicative bypass of an abasic DNA lesion by DNA polymerase III.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 22 ·1992-11-15 ·Pages 10777-81

Rajagopalan M, Lu C, Woodgate R, O'Donnell M, Goodman MF, Echols H

Abstract

The introduction of a replication-inhibiting lesion into the DNA of Escherichia coli generates the induced, multigene SOS response. One component of the SOS response is a marked increase in mutation rate, dependent on RecA protein and the induced mutagenesis proteins UmuC and UmuD. A variety of previous indirect approaches have indicated that SOS mutagenesis results from replicative bypass of the DNA lesion by DNA polymerase III (pol III) holoenzyme in a reaction mediated by RecA, UmuC, and a processed form of UmuD termed UmuD'. To study the biochemistry of SOS mutagenesis, we have reconstituted replicative bypass with a defined in vitro system containing purified protein and a DNA substrate with a single abasic DNA lesion. The replicative bypass reaction requires pol III, UmuC, UmuD', and RecA. The nonprocessed UmuD protein does not replace UmuD' but inhibits the bypass activity of UmuD', perhaps by sequestering UmuD' in a heterodimer. Our experiments demonstrate directly that the UmuC-UmuD' complex and RecA act to rescue an otherwise stalled pol III holoenzyme at a replication-blocking DNA lesion.

MeSH Terms
Bacterial Proteins/isolation & purification,metabolism DNA Damage DNA Polymerase III/metabolism DNA Replication DNA, Bacterial/biosynthesis,isolation & purification DNA-Directed DNA Polymerase Escherichia coli/genetics,metabolism Escherichia coli Proteins Kinetics Models, Structural Rec A Recombinases/isolation & purification,metabolism SOS Response, Genetics Templates, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins UmuC protein, E coli Rec A Recombinases DNA Polymerase III DNA-Directed DNA Polymerase UmuD protein, E coli
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Rajagopalan M
Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720.
Lu C
Woodgate R
O'Donnell M
Goodman M F
Echols H
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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.89.22.10777
Published
1992-11-15
Pages
10777-81
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50425
Subset
IM
Grants
NCI NIH HHS · CA41655 · United States
NIGMS NIH HHS · GM38839 · United States
NIGMS NIH HHS · GM42554 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.22.10777 PMC Full Text View journal details
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