Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 22 October 2004:
Vol. 306. no. 5696, pp. 683 - 685
DOI: 10.1126/science.1099824
Prev | Table of Contents | Next

Reports

Mycobacterial Ku and Ligase Proteins Constitute a Two-Component NHEJ Repair Machine

Marina Della,1*{dagger} Phillip L. Palmbos,2{dagger} Hui-Min Tseng,3{dagger} Louise M. Tonkin,1,4 James M. Daley,2 Leana M. Topper,2 Robert S. Pitcher,4 Alan E. Tomkinson,5 Thomas E. Wilson,2 Aidan J. Doherty4{ddagger}

In mammalian cells, repair of DNA double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ) is critical for genome stability. Although the end-bridging and ligation steps of NHEJ have been reconstituted in vitro, little is known about the end-processing reactions that occur before ligation. Recently, functionally homologous end-bridging and ligation activities have been identified in prokarya. Consistent with its homology to polymerases and nucleases, we demonstrate that DNA ligase D from Mycobacterium tuberculosis (Mt-Lig) possesses a unique variety of nucleotidyl transferase activities, including gap-filling polymerase, terminal transferase, and primase, and is also a 3' to 5' exonuclease. These enzyme activities allow the Mt-Ku and Mt-Lig proteins to join incompatible DSB ends in vitro, as well as to reconstitute NHEJ in vivo in yeast. These results demonstrate that prokaryotic Ku and ligase form a bona fide NHEJ system that encodes all the recognition, processing, and ligation activities required for DSB repair.

1 Cambridge Institute for Medical Research, University of Cambridge, Department of Haematology, Hills Road, Cambridge CB2 2XY, UK.
2 Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan 48109–0602, USA.
3 Molecular Medicine Graduate Program, Institute of Biotechnology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78245–3207, USA.
4 Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton BN1 9RQ, UK.
5 Department of Radiation Oncology and Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201–1559, USA.



* Present address: The Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, UK.

{dagger} These authors contributed equally to this work.

{ddagger} To whom correspondence should be addressed. E-mail: AJD21{at}sussex.ac.uk

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Chromatin Organization and Radio Resistance in the Bacterium Gemmata obscuriglobus.
A. Lieber, A. Leis, A. Kushmaro, A. Minsky, and O. Medalia (2009)
J. Bacteriol. 191, 1439-1445
   Abstract »    Full Text »    PDF »
Comparison of Responses to Double-Strand Breaks between Escherichia coli and Bacillus subtilis Reveals Different Requirements for SOS Induction.
L. A. Simmons, A. I. Goranov, H. Kobayashi, B. W. Davies, D. S. Yuan, A. D. Grossman, and G. C. Walker (2009)
J. Bacteriol. 191, 1152-1161
   Abstract »    Full Text »    PDF »
RecBCD Enzyme and the Repair of Double-Stranded DNA Breaks.
M. S. Dillingham and S. C. Kowalczykowski (2008)
Microbiol. Mol. Biol. Rev. 72, 642-671
   Abstract »    Full Text »    PDF »
Recruitment of Saccharomyces cerevisiae Dnl4-Lif1 Complex to a Double-Strand Break Requires Interactions With Yku80 and the Xrs2 FHA Domain.
P. L. Palmbos, D. Wu, J. M. Daley, and T. E. Wilson (2008)
Genetics 180, 1809-1819
   Abstract »    Full Text »    PDF »
Editing of misaligned 3'-termini by an intrinsic 3'-5' exonuclease activity residing in the PHP domain of a family X DNA polymerase.
B. Banos, J. M. Lazaro, L. Villar, M. Salas, and M. de Vega (2008)
Nucleic Acids Res. 36, 5736-5749
   Abstract »    Full Text »    PDF »
Bacterial Nonhomologous End Joining Ligases Preferentially Seal Breaks with a 3'-OH Monoribonucleotide.
H. Zhu and S. Shuman (2008)
J. Biol. Chem. 283, 8331-8339
   Abstract »    Full Text »    PDF »
Mechanistic flexibility as a conserved theme across 3 billion years of nonhomologous DNA end-joining.
J. Gu and M. R. Lieber (2008)
Genes & Dev. 22, 411-415
   Full Text »    PDF »
The pathways and outcomes of mycobacterial NHEJ depend on the structure of the broken DNA ends.
J. Aniukwu, M. S. Glickman, and S. Shuman (2008)
Genes & Dev. 22, 512-527
   Abstract »    Full Text »    PDF »
Structure of a NHEJ Polymerase-Mediated DNA Synaptic Complex.
N. C. Brissett, R. S. Pitcher, R. Juarez, A. J. Picher, A. J. Green, T. R. Dafforn, G. C. Fox, L. Blanco, and A. J. Doherty (2007)
Science 318, 456-459
   Abstract »    Full Text »    PDF »
Ku Heterodimer-Independent End Joining in Trypanosoma brucei Cell Extracts Relies upon Sequence Microhomology.
P. Burton, D. J. McBride, J. M. Wilkes, J. D. Barry, and R. McCulloch (2007)
Eukaryot. Cell 6, 1773-1781
   Abstract »    Full Text »    PDF »
Mycobacterial Nonhomologous End Joining Mediates Mutagenic Repair of Chromosomal Double-Strand DNA Breaks.
N. C. Stephanou, F. Gao, P. Bongiorno, S. Ehrt, D. Schnappinger, S. Shuman, and M. S. Glickman (2007)
J. Bacteriol. 189, 5237-5246
   Abstract »    Full Text »    PDF »
Characterization of Agrobacterium tumefaciens DNA ligases C and D.
H. Zhu and S. Shuman (2007)
Nucleic Acids Res. 35, 3631-3645
   Abstract »    Full Text »    PDF »
Mycobacterial UvrD1 Is a Ku-dependent DNA Helicase That Plays a Role in Multiple DNA Repair Events, Including Double-strand Break Repair.
K. M. Sinha, N. C. Stephanou, F. Gao, M. S. Glickman, and S. Shuman (2007)
J. Biol. Chem. 282, 15114-15125
   Abstract »    Full Text »    PDF »
Role of DNA Repair by Nonhomologous-End Joining in Bacillus subtilis Spore Resistance to Extreme Dryness, Mono- and Polychromatic UV, and Ionizing Radiation.
R. Moeller, E. Stackebrandt, G. Reitz, T. Berger, P. Rettberg, A. J. Doherty, G. Horneck, and W. L. Nicholson (2007)
J. Bacteriol. 189, 3306-3311
   Abstract »    Full Text »    PDF »
Nucleotide Misincorporation, 3'-Mismatch Extension, and Responses to Abasic Sites and DNA Adducts by the Polymerase Component of Bacterial DNA Ligase D.
L. Yakovleva and S. Shuman (2006)
J. Biol. Chem. 281, 25026-25040
   Abstract »    Full Text »    PDF »
Substrate Specificity and Structure-Function Analysis of the 3'-Phosphoesterase Component of the Bacterial NHEJ Protein, DNA Ligase D.
H. Zhu and S. Shuman (2006)
J. Biol. Chem. 281, 13873-13881
   Abstract »    Full Text »    PDF »
Crystal Structure and Nonhomologous End-joining Function of the Ligase Component of Mycobacterium DNA Ligase D.
D. Akey, A. Martins, J. Aniukwu, M. S. Glickman, S. Shuman, and J. M. Berger (2006)
J. Biol. Chem. 281, 13412-13423
   Abstract »    Full Text »    PDF »
Atomic structure and nonhomologous end-joining function of the polymerase component of bacterial DNA ligase D.
H. Zhu, J. Nandakumar, J. Aniukwu, L. K. Wang, M. S. Glickman, C. D. Lima, and S. Shuman (2006)
PNAS 103, 1711-1716
   Abstract »    Full Text »    PDF »
Mutations of the Yku80 C Terminus and Xrs2 FHA Domain Specifically Block Yeast Nonhomologous End Joining.
P. L. Palmbos, J. M. Daley, and T. E. Wilson (2005)
Mol. Cell. Biol. 25, 10782-10790
   Abstract »    Full Text »    PDF »
Essential Constituents of the 3'-Phosphoesterase Domain of Bacterial DNA Ligase D, a Nonhomologous End-joining Enzyme.
H. Zhu, L. K. Wang, and S. Shuman (2005)
J. Biol. Chem. 280, 33707-33715
   Abstract »    Full Text »    PDF »
DNA Joint Dependence of Pol X Family Polymerase Action in Nonhomologous End Joining.
J. M. Daley, R. L. V. Laan, A. Suresh, and T. E. Wilson (2005)
J. Biol. Chem. 280, 29030-29037
   Abstract »    Full Text »    PDF »
Origin and evolution of the archaeo-eukaryotic primase superfamily and related palm-domain proteins: structural insights and new members.
L. M. Iyer, E. V. Koonin, D. D. Leipe, and L. Aravind (2005)
Nucleic Acids Res. 33, 3875-3896
   Abstract »    Full Text »    PDF »
Novel 3'-Ribonuclease and 3'-Phosphatase Activities of the Bacterial Non-homologous End-joining Protein, DNA Ligase D.
H. Zhu and S. Shuman (2005)
J. Biol. Chem. 280, 25973-25981
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)