| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 275, Issue 1, 507-513, January 7, 2000
From the The RecBCD enzyme of Escherichia coli
is an ATP-dependent DNA exonuclease and a helicase. Its
exonuclease activity is subject to regulation by an octameric
nucleotide sequence called
A Single Nuclease Active Site of the Escherichia
coli RecBCD Enzyme Catalyzes Single-stranded DNA
Degradation in Both Directions*
,§, and¶
Molecular and Cell Biology Program and
¶ Department of Chemistry and Biochemistry, University of
Maryland, College Park, Maryland 20742
. In this study, site-directed mutations
were made in the carboxyl-terminal nuclease domain of the RecB subunit,
and their effects on RecBCD's enzymatic activities were investigated.
Mutation of two amino acid residues, Asp1067 and
Lys1082, abolished nuclease activity on both single- and
double-stranded DNA. Together with Asp1080, these residues
compose a motif that is similar to one shown to form the active site of
several restriction endonucleases. The nuclease reactions catalyzed by
the RecBCD enzyme should therefore follow the same mechanism as these
restriction endonucleases. Furthermore, the mutant enzymes were unable
to produce -specific fragments that are thought to result from the
3'-5' and 5'-3' single-stranded exonuclease activities of the enzyme
during its reaction with -containing double-stranded DNA. The
results show that the nuclease active site in the RecB C-terminal
30-kDa domain is the universal nuclease active site of RecBCD that is
responsible for DNA degradation in both directions during
the reaction with double-stranded DNA. A novel explanation for the
observed nuclease polarity switch and RecBCD-DNA interaction is offered.
*
This work was supported by National Institutes of Health
Grant GM39777.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Chemistry
and Biochemistry, University of Maryland, College Park, MD 20742. Tel.:
301-405-1821; Fax: 301-314-9121; E-mail: dj13@umail.umd.edu.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Ting, E. A. Kouzminova, and A. Kuzminov Synthetic Lethality with the dut Defect in Escherichia coli Reveals Layers of DNA Damage of Increasing Complexity Due to Uracil Incorporation J. Bacteriol., September 1, 2008; 190(17): 5841 - 5854. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Sisakova, L. K. Stanley, M. Weiserova, and M. D. Szczelkun A RecB-family nuclease motif in the Type I restriction endonuclease EcoR124I Nucleic Acids Res., July 1, 2008; 36(12): 3939 - 3949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Hill, T. Woodward, A. Reger, R. Baker, and T. Dinse Role for the RecBCD Recombination Pathway for pilE Gene Variation in Repair-Proficient Neisseria gonorrhoeae J. Bacteriol., November 15, 2007; 189(22): 7983 - 7990. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Ivancic-Bace, I. Vlasic, G. Cogelja-Cajo, K. Brcic-Kostic, and E. Salaj-Smic Roles of PriA Protein and Double-Strand DNA Break Repair Functions in UV-Induced Restriction Alleviation in Escherichia coli Genetics, December 1, 2006; 174(4): 2137 - 2149. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Chedin, N. Handa, M. S. Dillingham, and S. C. Kowalczykowski The AddAB Helicase/Nuclease Forms a Stable Complex with Its Cognate {chi} Sequence During Translocation J. Biol. Chem., July 7, 2006; 281(27): 18610 - 18617. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Ivancic-Bace, I. Vlasic, E. Salaj-Smic, and K. Brcic-Kostic Genetic Evidence for the Requirement of RecA Loading Activity in SOS Induction after UV Irradiation in Escherichia coli. J. Bacteriol., July 1, 2006; 188(14): 5024 - 5032. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Thermic Functions of Multiple Exonucleases Are Essential for Cell Viability, DNA Repair and Homologous Recombination in recD Mutants of Escherichia coli Genetics, April 1, 2006; 172(4): 2057 - 2069. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Ivancic-Bace, E. Salaj-Smic, and K. Brcic-Kostic Effects of recJ, recQ, and recFOR Mutations on Recombination in Nuclease-Deficient recB recD Double Mutants of Escherichia coli J. Bacteriol., February 15, 2005; 187(4): 1350 - 1356. [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. Miranda and A. Kuzminov Chromosomal Lesion Suppression and Removal in Escherichia coli via Linear DNA Degradation Genetics, April 1, 2003; 163(4): 1255 - 1271. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Ivancic-Bace, P. Peharec, S. Moslavac, N. Skrobot, E. Salaj-Smic, and K. Brcic-Kostic RecFOR Function Is Required for DNA Repair and Recombination in a RecA Loading-Deficient recB Mutant of Escherichia coli Genetics, February 1, 2003; 163(2): 485 - 494. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. K. Amundsen, A. F. Taylor, and G. R. Smith A Domain of RecC Required for Assembly of the Regulatory RecD Subunit Into the Escherichia coli RecBCD Holoenzyme Genetics, June 1, 2002; 161(2): 483 - 492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Wang, I. B. Castano, C. Adams, C. Vu, D. Fitzhugh, and M. F. Christman Structure/Function Analysis of the Saccharomyces cerevisiae Trf4/Pol {sigma} DNA Polymerase Genetics, February 1, 2002; 160(2): 381 - 391. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. S. Makarova, L. Aravind, N. V. Grishin, I. B. Rogozin, and E. V. Koonin A DNA repair system specific for thermophilic Archaea and bacteria predicted by genomic context analysis Nucleic Acids Res., January 15, 2002; 30(2): 482 - 496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. W. Chang and D. A. Julin Structure and Function of the Escherichia coli RecE Protein, a Member of the RecB Nuclease Domain Family J. Biol. Chem., November 30, 2001; 276(49): 46004 - 46010. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Quiberoni, I. Biswas, M. El Karoui, L. Rezaiki, P. Tailliez, and A. Gruss In Vivo Evidence for Two Active Nuclease Motifs in the Double-Strand Break Repair Enzyme RexAB of Lactococcus lactis J. Bacteriol., July 1, 2001; 183(13): 4071 - 4078. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. M. Ellis, D. Yu, T. DiTizio, and D. L. Court High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides PNAS, May 24, 2001; (2001) 121164898. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Daiyasu, K. Komori, S. Sakae, Y. Ishino, and H. Toh Hjc resolvase is a distantly related member of the type II restriction endonuclease family Nucleic Acids Res., November 15, 2000; 28(22): 4540 - 4543. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Aravind, K. S. Makarova, and E. V. Koonin SURVEY AND SUMMARY: Holliday junction resolvases and related nucleases: identification of new families, phyletic distribution and evolutionary trajectories Nucleic Acids Res., September 15, 2000; 28(18): 3417 - 3432. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Li, R. Geyer, and W. Tan Using molecular beacons as a sensitive fluorescence assay for enzymatic cleavage of single-stranded DNA Nucleic Acids Res., June 1, 2000; 28(11): e52 - e52. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. M. Ellis, D. Yu, T. DiTizio, and D. L. Court High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides PNAS, June 5, 2001; 98(12): 6742 - 6746. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
