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UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops.

Publication ,  Journal Article
Dudas, KC; Kreuzer, KN
Published in: Mol Cell Biol
April 2001

The UvsW protein of bacteriophage T4 is involved in many aspects of phage DNA metabolism, including repair, recombination, and recombination-dependent replication. UvsW has also been implicated in the repression of origin-dependent replication at late times of infection, when UvsW is normally synthesized. Two well-characterized T4 origins, ori(uvsY) and ori(34), are believed to initiate replication through an R-loop mechanism. Here we provide both in vivo and in vitro evidence that UvsW is an RNA-DNA helicase that catalyzes the dissociation of RNA from origin R-loops. Two-dimensional gel analyses show that the replicative intermediates formed at ori(uvsY) persist longer in a uvsW mutant infection than in a wild-type infection. In addition, the inappropriate early expression of UvsW protein results in the loss of these replicative intermediates. Using a synthetic origin R-loop, we also demonstrate that purified UvsW functions as a helicase that efficiently dissociates RNA from R-loops. These and previous results from a number of studies provide strong evidence that UvsW is a molecular switch that allows T4 replication to progress from a mode that initiates from R-loops at origins to a mode that initiates from D-loops formed by recombination proteins.

Duke Scholars

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

April 2001

Volume

21

Issue

8

Start / End Page

2706 / 2715

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Replication Origin
  • RNA, Viral
  • Nucleic Acid Conformation
  • Mutation
  • Genes, Viral
  • Escherichia coli
  • Developmental Biology
  • DNA Replication
 

Citation

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Dudas, K. C., & Kreuzer, K. N. (2001). UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops. Mol Cell Biol, 21(8), 2706–2715. https://doi.org/10.1128/MCB.21.8.2706-2715.2001
Dudas, K. C., and K. N. Kreuzer. “UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops.Mol Cell Biol 21, no. 8 (April 2001): 2706–15. https://doi.org/10.1128/MCB.21.8.2706-2715.2001.
Dudas KC, Kreuzer KN. UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops. Mol Cell Biol. 2001 Apr;21(8):2706–15.
Dudas, K. C., and K. N. Kreuzer. “UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops.Mol Cell Biol, vol. 21, no. 8, Apr. 2001, pp. 2706–15. Pubmed, doi:10.1128/MCB.21.8.2706-2715.2001.
Dudas KC, Kreuzer KN. UvsW protein regulates bacteriophage T4 origin-dependent replication by unwinding R-loops. Mol Cell Biol. 2001 Apr;21(8):2706–2715.

Published In

Mol Cell Biol

DOI

ISSN

0270-7306

Publication Date

April 2001

Volume

21

Issue

8

Start / End Page

2706 / 2715

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Replication Origin
  • RNA, Viral
  • Nucleic Acid Conformation
  • Mutation
  • Genes, Viral
  • Escherichia coli
  • Developmental Biology
  • DNA Replication