|
Volume 270,
Number 41,
Issue of October 13, 1995 pp. 24459-24467
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Strand
Specificity of Nicking of DNA at Chi Sites by RecBCD Enzyme
MODULATION BY ATP AND MAGNESIUM LEVELS
(Received for publication, May 15, 1995)
Andrew F.
Taylor,
Gerald
R.
Smith
RecBCD enzyme is essential for the major pathway of homologous
recombination of linear DNA in Escherichia coli. It is a
potent nuclease and helicase and, during its unwinding of
double-stranded DNA, makes single-strand scissions in the vicinity of
Chi recombination hot spots. We report here that both the strand that
is cut and the position of the cuts relative to Chi depended on the ATP
to Mg ratio. With ATP in excess, Chi-dependent nicks
occurred, as we have previously reported, four to six nucleotides to
the 3`-side of the Chi octamer (5`-GCTGGTGG-3`) and were detected only
on the strand bearing that sequence. Three differences were seen with
Mg in excess. 1) Chi-dependent 3`-ends were produced
on the GCTGGTGG-containing strand closer to and within the Chi octamer.
2) Chi-dependent cuts occurred on the complementary DNA strand. 3)
RecBCD enzyme destroyed the 3`-terminated strand of DNA from its entry
point up to the vicinity of the Chi site, as others have previously
reported. We show here that, with Mg in excess, the
enzyme continued to travel along DNA, after encountering a Chi site,
releasing both strands of the DNA distal to Chi as single strands. We
discuss potential biological consequences of these two modes of RecBCD
enzyme-Chi interaction.

CiteULike Complore Connotea Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
M. S. Dillingham and S. C. Kowalczykowski
RecBCD Enzyme and the Repair of Double-Stranded DNA Breaks
Microbiol. Mol. Biol. Rev.,
December 1, 2008;
72(4):
642 - 671.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. K. Amundsen, A. F. Taylor, M. Reddy, and G. R. Smith
Intersubunit signaling in RecBCD enzyme, a complex protein machine regulated by Chi hot spots
Genes & Dev.,
December 15, 2007;
21(24):
3296 - 3307.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. K. Amundsen and G. R. Smith
Chi Hotspot Activity in Escherichia coli Without RecBCD Exonuclease Activity: Implications for the Mechanism of Recombination
Genetics,
January 1, 2007;
175(1):
41 - 54.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Spies, M. S. Dillingham, and S. C. Kowalczykowski
Translocation by the RecB Motor Is an Absolute Requirement for {chi}-Recognition and RecA Protein Loading by RecBCD Enzyme
J. Biol. Chem.,
November 4, 2005;
280(44):
37078 - 37087.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Kulkarni and D. A. Julin
Specific inhibition of the E.coli RecBCD enzyme by Chi sequences in single-stranded oligodeoxyribonucleotides
Nucleic Acids Res.,
July 14, 2004;
32(12):
3672 - 3682.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. M. Osborn, F. M. da Silva Tatley, L. M. Steyn, R. W. Pickup, and J. R. Saunders
Mosaic plasmids and mosaic replicons: evolutionary lessons from the analysis of genetic diversity in IncFII-related replicons
Microbiology,
September 1, 2000;
146(9):
2267 - 2275.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. Wang, R. Chen, and D. A. Julin
A Single Nuclease Active Site of the Escherichia coli RecBCD Enzyme Catalyzes Single-stranded DNA Degradation in Both Directions
J. Biol. Chem.,
January 7, 2000;
275(1):
507 - 513.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Kuzminov
Recombinational Repair of DNA Damage in Escherichia coli and Bacteriophage lambda
Microbiol. Mol. Biol. Rev.,
December 1, 1999;
63(4):
751 - 813.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. R. Motamedi, S. K. Szigety, and S. M. Rosenberg
Double-strand-break repair recombination in Escherichia coli: physical evidence for a DNA replication mechanism in vivo
Genes & Dev.,
November 1, 1999;
13(21):
2889 - 2903.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
A. F. Taylor and G. R. Smith
Regulation of homologous recombination: Chi inactivates RecBCD enzyme by disassembly of the three subunits
Genes & Dev.,
April 1, 1999;
13(7):
890 - 900.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
A. Kuzminov and F. W. Stahl
Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication
Genes & Dev.,
February 1, 1999;
13(3):
345 - 356.
[Abstract]
[Full Text]
|
 |
|

|
 |

|
 |
 
M. K. B. Berlyn
Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map
Microbiol. Mol. Biol. Rev.,
September 1, 1998;
62(3):
814 - 984.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Friedman-Ohana and A. Cohen
Heteroduplex joint formation in Escherichia coli recombination is initiated by pairing of a 3'-ending strand
PNAS,
June 9, 1998;
95(12):
6909 - 6914.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. Noirot and R. D. Kolodner
DNA Strand Invasion Promoted by Escherichia coli RecT Protein
J. Biol. Chem.,
May 15, 1998;
273(20):
12274 - 12280.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
R. Friedman-Ohana, I. Karunker, and A. Cohen
Chi-Dependent Intramolecular Recombination in Escherichia coli
Genetics,
February 1, 1998;
148(2):
545 - 558.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
H.-W. Chen, B. Ruan, M. Yu, J.-d. Wang, and D. A. Julin
The RecD Subunit of the RecBCD Enzyme from Escherichia coli Is a Single-stranded DNA-dependent ATPase
J. Biol. Chem.,
April 11, 1997;
272(15):
10072 - 10079.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D G Anderson and S C Kowalczykowski
The recombination hot spot chi is a regulatory element that switches the polarity of DNA degradation by the RecBCD enzyme.
Genes & Dev.,
March 1, 1997;
11(5):
571 - 581.
[Abstract]
[PDF]
|
 |
|

|
 |

|
 |
 
A. F. Taylor and G. R. Smith
Monomeric RecBCD Enzyme Binds and Unwinds DNA
J. Biol. Chem.,
October 13, 1995;
270(41):
24451 - 24458.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|