phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 72 ·No. 10 ·1975-10-00 ·Pages 3907-11

Schuster H, Mikolajczyk M, Rohrschneider J, Geschke B

Abstract

Ammonium sulfate fractionation of crude extracts of E. coli yields a soluble enzyme fraction (about 25-fold purification) that catalyzes the conversion of phiX174 single-stranded DNA to duplex DNA. The reaction is rifampicin-resistant, requires single-stranded DNA, Mg++, deoxynucleoside triphosphates, and ATP, and is stimulated by KCl. Such soluble enzyme fractions were prepared from E. coli strains carrying the prophage mutant P1bac, in which the viral dnaB analog (ban) protein is expressed constitutively, or P1bacban, in which the expression of ban protein is prevented. DNA-synthesizing activity of ban protein containing fractions from wild-type or dnaB(P1bac) lysogens was more temperature-resistant than that from E. coli containing only wild-type dnaB protein, whereas that from dnaB(P1bacban) lysogens of dnaB cells was extremely thermolabile. It is suggested that the temperature-resistant DNA synthesis with fractions from P1bac lysogens is mediated by the P1 ban protein.

MeSH Terms

Bacterial Proteins/metabolism Coliphages/drug effects,metabolism DNA Replication/drug effects DNA, Single-Stranded/metabolism DNA, Viral/metabolism Escherichia coli/drug effects,metabolism Hydrogen-Ion Concentration Kinetics Magnesium/pharmacology Mutation Potassium Chloride/pharmacology Rifampin/pharmacology Streptomycin Temperature

Chemicals

Bacterial Proteins DNA, Single-Stranded DNA, Viral Potassium Chloride Magnesium Rifampin Streptomycin

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Schuster H

Mikolajczyk M

Rohrschneider J

Geschke B

References (15)

15 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.72.10.3907

Published

1975-10-00

Pages

3907-11

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC433105

Subset

External Links

PubMed source DOI: 10.1073/pnas.72.10.3907 PMC Full Text View journal details

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phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.

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