The Escherichia coli dnaB replication protein is a DNA helicase

J. H. LeBowitz, R. McMacken

Research output: Contribution to journalArticlepeer-review

Abstract

Genetic and biochemical analyses indicate that the Escherichia coli dnaB replication protein functions in the propagation of replication forks in the bacterial chromosome. We have found that the dnaB protein is a DNA helicase that is capable of unwinding extensive stretches of double-stranded DNA. We constructed a partially duplex DNA substrate, containing two preformed forks of single-stranded DNA, which was used to characterize this helicase activity. The dnaB helicase (i) depends on the presence of a hydrolyzable ribonucleoside triphosphate, (ii) is maximally stimulated by a combination of E. coli single-stranded DNA-binding protein and E. coli primase, (iii) is inhibited by antibody directed against dnaB protein, and (iv) is inhibited by prior coating of the single-stranded regions of the helicase substrate with the E. coli single-stranded DNA-binding protein. It was determined that the dnaB protein moves 5' to 3' along single-stranded DNA, apparently in a processive fashion. To invade the duplex portion of the helicase substrate, the dnaB protein requires a 3'-terminal extension of single-stranded DNA in the strand to which it is not bound. Under optimal conditions at 30°C, greater than 1 kilobase pair of duplex DNA can be unwound within 30 s. Based on these findings and other available data, we propose that the dnaB protein is the primary replicative helicase of E. coli and that it actively and processively migrates along the lagging strand template, serving both to unwind the DNA duplex in advance of the leading straqnd and to potentiate synthesis by the bacterial primase of RNA primers for the nascent (Okazaki) fragments of the lagging strand.

Original languageEnglish (US)
Pages (from-to)4738-4748
Number of pages11
JournalJournal of Biological Chemistry
Volume261
Issue number10
StatePublished - 1986

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'The Escherichia coli dnaB replication protein is a DNA helicase'. Together they form a unique fingerprint.

Cite this