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PMID: 9009281 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

ATP-dependent resolution of R-loops at the ColE1 replication origin by Escherichia coli RecG protein, a Holliday junction-specific helicase.

The EMBO journal ·Vol. 16 ·No. 1 ·1997-01-02 ·Pages 203-9

Fukuoh A, Iwasaki H, Ishioka K, Shinagawa H

Abstract

The RecG protein of Escherichia coli is a DNA helicase that promotes branch migration of the Holliday junctions. We found that overproduction of RecG protein drastically decreased copy numbers of ColE1-type plasmids, which require R-loop formation between the template DNA and a primer RNA transcript (RNA II) for the initiation of replication. RecG efficiently inhibited in vitro ColE1 DNA synthesis in a reconstituted system containing RNA polymerase, RNase HI and DNA polymerase I. RecG promoted dissociation of RNA II from the R-loop in a manner that required ATP hydrolysis. These results suggest that overproduced RecG inhibits the initiation of replication by prematurely resolving the R-loops formed at the replication origin region of these plasmids with its unique helicase activity. The possibility that RecG regulates the initiation of a unique mode of DNA replication, oriC-independent constitutive stable DNA replication, by its activity in resolving R-loops is discussed.

MeSH Terms
Adenosine Triphosphate/metabolism Bacterial Proteins/genetics,physiology Bacteriocin Plasmids/genetics DNA Helicases/genetics,physiology DNA Replication/physiology DNA, Bacterial/biosynthesis,chemistry,metabolism Escherichia coli/genetics Escherichia coli Proteins Gene Dosage Hydrolysis Nucleic Acid Conformation RNA, Bacterial/metabolism Replication Origin
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins RNA, Bacterial RecG protein, E coli Adenosine Triphosphate DNA Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Fukuoh A
Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
Iwasaki H
Ishioka K
Shinagawa H
References (22)
22 references, click to expand
  1. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H.
    Proc Natl Acad Sci U S A. 1980 May;77(5):2450-4 PMID: 6156450
  2. Recombination by replication.
    Cell. 1996 May 31;85(5):625-7 PMID: 8646771
  3. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  4. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library.
    Cell. 1987 Jul 31;50(3):495-508 PMID: 3038334
  5. Use of T7 RNA polymerase to direct expression of cloned genes.
    Methods Enzymol. 1990;185:60-89 PMID: 2199796
  6. Evidence of abortive recombination in ruv mutants of Escherichia coli K12.
    Mol Gen Genet. 1991 Feb;225(2):266-72 PMID: 2005868
  7. Conjugational recombination in resolvase-deficient ruvC mutants of Escherichia coli K-12 depends on recG.
    J Bacteriol. 1991 Sep;173(17):5414-8 PMID: 1653210
  8. Molecular organization and nucleotide sequence of the recG locus of Escherichia coli K-12.
    J Bacteriol. 1991 Nov;173(21):6837-43 PMID: 1938888
  9. Escherichia coli RuvC protein is an endonuclease that resolves the Holliday structure.
    EMBO J. 1991 Dec;10(13):4381-9 PMID: 1661673
  10. Formation and resolution of recombination intermediates by E. coli RecA and RuvC proteins.
    Nature. 1991 Dec 19-26;354(6354):506-10 PMID: 1758493
  11. The nucleotide sequence of recG, the distal spo operon gene in Escherichia coli K-12.
    Gene. 1992 Jan 2;110(1):95-9 PMID: 1544582
  12. Interaction of Escherichia coli RuvA and RuvB proteins with synthetic Holliday junctions.
    Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5452-6 PMID: 1608954
  13. Escherichia coli RuvA and RuvB proteins specifically interact with Holliday junctions and promote branch migration.
    Genes Dev. 1992 Nov;6(11):2214-20 PMID: 1427081
  14. Dissociation of synthetic Holliday junctions by E. coli RecG protein.
    EMBO J. 1993 Jan;12(1):17-22 PMID: 8428576
  15. Formation of a RuvAB-Holliday junction complex in vitro.
    J Mol Biol. 1993 Jul 20;232(2):397-405 PMID: 8393934
  16. Reverse branch migration of Holliday junctions by RecG protein: a new mechanism for resolution of intermediates in recombination and DNA repair.
    Cell. 1993 Oct 22;75(2):341-50 PMID: 8402917
  17. RecA, Tus protein and constitutive stable DNA replication in Escherichia coli rnhA mutants.
    Mol Gen Genet. 1994 Sep 1;244(5):557-62 PMID: 8078483
  18. Branch migration of Holliday junctions: identification of RecG protein as a junction specific DNA helicase.
    EMBO J. 1994 Nov 1;13(21):5220-8 PMID: 7957087
  19. Roles of ruvA, ruvC and recG gene functions in normal and DNA damage-inducible replication of the Escherichia coli chromosome.
    Genetics. 1994 Aug;137(4):895-902 PMID: 7982571
  20. Escherichia coli RecG and RecA proteins in R-loop formation.
    EMBO J. 1995 May 15;14(10):2385-92 PMID: 7774596
  21. Branch migration of three-strand recombination intermediates by RecG, a possible pathway for securing exchanges initiated by 3'-tailed duplex DNA.
    EMBO J. 1995 Jul 17;14(14):3302-10 PMID: 7628432
  22. Effects of point mutations on formation and structure of the RNA primer for ColE1 DNA replication.
    Cell. 1984 Feb;36(2):513-22 PMID: 6198094
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.1.203
Published
1997-01-02
Pages
203-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1169627
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/16.1.203 PMC Full Text View journal details
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