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Science 22 October 2004:
Vol. 306. no. 5696, pp. 683 - 685
DOI: 10.1126/science.1099824
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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

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DNA Joint Dependence of Pol X Family Polymerase Action in Nonhomologous End Joining.
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