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PMID: 127175 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Association of phiX174 DNA-dependent ATPase activity with an Escherichia coli protein, replication factor Y, required for in vitro synthesis of phiX174 DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 72 ·No. 9 ·1975-09-00 ·Pages 3342-6

Wickner S, Hurwitz J

Abstract

phiX174 DNA-dependent DNA synthesis is catalyzed in vitro by the combination of at least 11 purified protein fractions: dnaB, dnaC(D), and dnaG gene products, DNA polymerase III, DNA elongation factors I and II, DNA binding protein, and replication factors W, X, Y, and Z. The reaction requires ATP, 4 dNTPs, and Mg+2 and is specific for phiX174 (or phiXahb) DNA. Purified replication factor Y contains phiX174 (or phiXahb) DNA-dependent ATPase (or dATPase) activity. The ATPase activity is poorly stimulated by other single-stranded DNA, by double-stranded DNA, or by RNA. The products of the phiX174 DNA-dependent ATPase activity of factor Y are Pi and ADP (or dADP). The association of phiX174 DNA-dependent ATPase activity with factor Y was shown in the following ways: (a) the two activities copurified with a constant ratio; (b) they comigrated on native polyacrylamide gel electrophoresis; (c) both activities were heat-inactivated at the same rate; and (d) both showed identical patterns of N-ethylmaleimide sensitivity.

MeSH Terms
Adenosine Triphosphatases/analysis,antagonists & inhibitors,metabolism Cell-Free System Coliphages/metabolism DNA Replication DNA, Viral/biosynthesis,pharmacology Enzyme Activation Escherichia coli/enzymology Ethylmaleimide/pharmacology Hot Temperature
Chemicals
DNA, Viral Adenosine Triphosphatases Ethylmaleimide
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wickner S
Hurwitz J
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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.72.9.3342
Published
1975-09-00
Pages
3342-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC432988
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.72.9.3342 PMC Full Text View journal details
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