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PMID: 4197093 Published · ppublish English Journal Article

Studies on in vitro DNA synthesis. Purification of the dna G gene product from Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 70 ·No. 5 ·1973-05-00 ·Pages 1613-8

Wickner S, Wright M, Hurwitz J

Abstract

varphiX174 DNA-dependent dNMP incorporation is temperature-sensitive (ts) in extracts of uninfected E. coli dna A, B, C, D, E, and G ts strains. DNA synthesis can be restored in heat-inactivated extracts of various dna ts mutants by addition of extracts of wild-type or other dna ts mutants. A protein that restores activity to heat-inactivated extracts of dna G ts cells has been extensively purified. This protein has also been purified from dna G ts cells and is thermolabile when compared to the wild-type protein. The purified dna G protein has a molecular weight of about 60,000, is insensitive to N-ethylmaleimide, and binds poorly to DNA. It does not stimulate heat-inactivated crude extracts of dna B, C, D, or E ts cells and lacks detectable RNA and DNA polymerase activities.

MeSH Terms
Ammonium Sulfate Bacterial Proteins/isolation & purification Binding Sites Centrifugation, Density Gradient Chromatography, DEAE-Cellulose Chromatography, Gel DNA Replication DNA, Bacterial/biosynthesis Dithiothreitol Edetic Acid Escherichia coli/growth & development,metabolism Freezing Genes Genetic Complementation Test Glycerol Methods Mutation Streptomycin Sulfates Temperature
Chemicals
Bacterial Proteins DNA, Bacterial Sulfates Edetic Acid Glycerol Ammonium Sulfate Dithiothreitol Streptomycin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wickner S
Wright M
Hurwitz J
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20 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.70.5.1613
Published
1973-05-00
Pages
1613-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC433553
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.70.5.1613 PMC Full Text View journal details
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