The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system

M. Zylicz, J. H. LeBowitz, Roger McMacken, C. Georgopoulos

Research output: Contribution to journalArticle

Abstract

The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.

Original languageEnglish (US)
Pages (from-to)6431-6435
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume80
Issue number21 I
StatePublished - 1983
Externally publishedYes

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Escherichia coli Proteins
DNA Replication
Adenosine Triphosphatases
Proteins
DNA
In Vitro Techniques
Escherichia coli
Mutation
Bacteriophages
Adenosine Diphosphate
Genes
Immunoglobulins

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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title = "The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system",
abstract = "The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.",
author = "M. Zylicz and LeBowitz, {J. H.} and Roger McMacken and C. Georgopoulos",
year = "1983",
language = "English (US)",
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journal = "Proceedings of the National Academy of Sciences of the United States of America",
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TY - JOUR

T1 - The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system

AU - Zylicz, M.

AU - LeBowitz, J. H.

AU - McMacken, Roger

AU - Georgopoulos, C.

PY - 1983

Y1 - 1983

N2 - The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.

AB - The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.

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