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PMID: 6280177 Published · ppublish English Journal Article

Purification and properties of the uvrA protein from Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 4 ·1982-02-00 ·Pages 988-92

Seeberg E, Steinum AL

Abstract

The uvrA+ gene product from Escherichia coli was purified to apparent homogeneity; the assay measured its ability to restore repair endonuclease activity in extracts from uvrA mutated cells. The uvrA protein is a 115,000 molecular weight DNA-binding protein having higher affinity for single-stranded than double-stranded DNA. It does not introduce single-strand breaks or alkali-labile bonds in native or UV-irradiated DNA, but it catalyzes hydrolysis of ATP to ADP and Pi. The ATPase activity is not DNA dependent and has a Km of 0.23 mM, which corresponds to the Km for the ATP requirement of the UV-endonuclease reaction catalyzed by the combined uvrA+, uvrB+, and uvrC+ gene products. ADP and adenosine 5'-[gamma-thio]triphosphate both inhibit the uvrA ATPase as well as the uvrABC endonuclease and also prevent specific binding of the uvrA proteins to UV-irradiated DNA. These results indicate that both the DNA-binding property and the ATPase activity of the uvrA protein are essential for uvrABC endonuclease activity and that the ATP requirement of the endonuclease reaction is determined by uvrA ATPase.

MeSH Terms
Adenosine Triphosphatases/isolation & purification,metabolism Adenosine Triphosphate/metabolism Bacterial Proteins/isolation & purification,metabolism,physiology Carrier Proteins/isolation & purification,metabolism DNA Repair DNA, Single-Stranded/metabolism DNA-Binding Proteins Endonucleases/metabolism Escherichia coli/analysis,metabolism Ultraviolet Rays
Chemicals
Bacterial Proteins Carrier Proteins DNA, Single-Stranded DNA-Binding Proteins Adenosine Triphosphate Endonucleases Adenosine Triphosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Seeberg E
Steinum A L
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24 references, click to expand
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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.79.4.988
Published
1982-02-00
Pages
988-92
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC345884
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.4.988 PMC Full Text View journal details
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