Mechanism of nonhomologous end-joining in mycobacteria: a low-fidelity repair system driven by ku, ligase d and ligase c

ABSTRACT DNA double-strand breaks (DSBs) can be repaired either via homologous recombination (HR) or nonhomologous end-joining (NHEJ). Both pathways are operative in eukaryotes, but bacteria

had been thought to rely on HR alone. Here we provide direct evidence that mycobacteria have a robust NHEJ pathway that requires Ku and a specialized polyfunctional ATP-dependent DNA ligase

(LigD). NHEJ of blunt-end and complementary 5′-overhang DSBs is highly mutagenic (∼50% error rate). Analysis of the recombination junctions ensuing from individual NHEJ events highlighted

the participation of several DNA end-remodeling activities, including template-dependent fill-in of 5′ overhangs, nontemplated addition of single nucleotides at blunt ends, and nucleolytic

resection. LigD itself has the template-dependent and template-independent polymerase functions _in vitro_ that compose the molecular signatures of NHEJ _in vivo_. Another ATP-dependent DNA

ligase (LigC) provides a backup mechanism for LigD-independent error-prone repair of blunt-end DSBs. We speculate that NHEJ allows mycobacteria to evade genotoxic host defense. Access

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SIMILAR CONTENT BEING VIEWED BY OTHERS POLΛ PROMOTES MICROHOMOLOGY-MEDIATED END-JOINING Article 19 December 2022 THE IMPORTANCE OF DNAPKCS FOR BLUNT DNA END JOINING IS MAGNIFIED WHEN XLF IS

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ACKNOWLEDGEMENTS This research was supported by US National Institutes of Health grants AI53417 (to M.S.G.) and GM63611 (to S.S.). M.S.G. is the recipient of research awards from the Ellison

Medical Foundation and the New York Academy of Medicine Speakers Fund for Biomedical Research. S.S. is an American Cancer Society Research Professor. AUTHOR INFORMATION Author notes *

Chunling Gong and Paola Bongiorno: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Immunology and Molecular Biology Programs, Sloan-Kettering Institute, New York,

10021, New York, USA Chunling Gong, Paola Bongiorno, Alexandra Martins, Nicolas C Stephanou, Hui Zhu, Stewart Shuman & Michael S Glickman * Division of Infectious Diseases, Memorial

Sloan Kettering Cancer Center, New York, 10021, New York, USA Chunling Gong, Paola Bongiorno, Alexandra Martins, Nicolas C Stephanou, Hui Zhu, Stewart Shuman & Michael S Glickman Authors

* Chunling Gong View author publications You can also search for this author inPubMed Google Scholar * Paola Bongiorno View author publications You can also search for this author inPubMed 

Google Scholar * Alexandra Martins View author publications You can also search for this author inPubMed Google Scholar * Nicolas C Stephanou View author publications You can also search for

this author inPubMed Google Scholar * Hui Zhu View author publications You can also search for this author inPubMed Google Scholar * Stewart Shuman View author publications You can also

search for this author inPubMed Google Scholar * Michael S Glickman View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHORS Correspondence

to Stewart Shuman or Michael S Glickman. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY TABLE 1

_Mycobacterium smegmatis_ deletion strains. (PDF 19 kb) SUPPLEMENTARY METHODS (PDF 89 KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Gong, C.,

Bongiorno, P., Martins, A. _et al._ Mechanism of nonhomologous end-joining in mycobacteria: a low-fidelity repair system driven by Ku, ligase D and ligase C. _Nat Struct Mol Biol_ 12,

304–312 (2005). https://doi.org/10.1038/nsmb915 Download citation * Received: 03 February 2005 * Accepted: 01 March 2005 * Published: 20 March 2005 * Issue Date: 01 April 2005 * DOI:

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