The neurodegenerative disease protein aprataxin resolves abortive dna ligation intermediates

ABSTRACT Ataxia oculomotor apraxia-1 (AOA1) is a neurological disorder caused by mutations in the gene (_APTX_) encoding aprataxin1,2. Aprataxin is a member of the histidine triad (HIT)

family of nucleotide hydrolases and transferases3, and inactivating mutations are largely confined to this HIT domain. Aprataxin associates with the DNA repair proteins XRCC1 and XRCC4,

which are partners of DNA ligase III and ligase IV, respectively4,5,6,7, suggestive of a role in DNA repair. Consistent with this, _APTX_-defective cell lines are sensitive to agents that

cause single-strand breaks and exhibit an increased incidence of induced chromosomal aberrations4,5,8. It is not, however, known whether aprataxin has a direct or indirect role in DNA

repair, or what the physiological substrate of aprataxin might be. Here we show, using purified aprataxin protein and extracts derived from either _APTX_-defective chicken DT40 cells or

_Aptx_-/- mouse primary neural cells, that aprataxin resolves abortive DNA ligation intermediates. Specifically, aprataxin catalyses the nucleophilic release of adenylate groups covalently

linked to 5′-phosphate termini at single-strand nicks and gaps, resulting in the production of 5′-phosphate termini that can be efficiently rejoined. These data indicate that neurological

disorders associated with _APTX_ mutations may be caused by the gradual accumulation of unrepaired DNA strand breaks resulting from abortive DNA ligation events. Access through your

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(2005) Article  ADS  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank D. Barnes and T. Lindahl for discussions and the gift of DNA ligase III–XRCC1 complex, and A. Ciccia

for advice and comments. We thank S. Rulten for the gift of recombinant His-tagged aprataxin, and M. Taylor for provision of AOA1 lymphoblastoid cells. This work was supported by Cancer

Research UK (S.C.W.), the EU DNA Repair Consortium (S.C.W.), the Medical Research Council (K.W.C.) and the NIH (P.J.M.). I.A. is supported by a fellowship from the European Molecular Biology

Organisation. Author Contributions I.A. made the initial discovery of DNA–adenylate activity, and I.A. and U.R. were responsible for experimental design and the generation of most of the

experimental data. S.F.E.-K., S.K. and P.M.C. were responsible for the development of vertebrate models and performed some of the experiments. P.J.M. and K.W.C. provided critical resources

and expertise, and contributed to writing the manuscript. S.C.W. was the project leader and produced the final version of the manuscript. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Cancer

Research UK, London Research Institute, Clare Hall Laboratories, South Mimms, Herts, EN6 3LD, UK Ivan Ahel, Ulrich Rass & Stephen C. West * Genome Damage and Stability Centre,

University of Sussex, BN1 9RQ, Falmer, Brighton, UK Sherif F. El-Khamisy, Paula M. Clements & Keith W. Caldecott * Biochemistry Department, Faculty of Pharmacy, Ain Shams University, PO

Box 11566, Cairo, Egypt Sherif F. El-Khamisy * Department of Genetics and Tumor Cell Biology, St. Jude Children's Research Hospital, 332N Lauderdale, Tennessee, 38105, Memphis, USA

Sachin Katyal & Peter J. McKinnon Authors * Ivan Ahel View author publications You can also search for this author inPubMed Google Scholar * Ulrich Rass View author publications You can

also search for this author inPubMed Google Scholar * Sherif F. El-Khamisy View author publications You can also search for this author inPubMed Google Scholar * Sachin Katyal View author

publications You can also search for this author inPubMed Google Scholar * Paula M. Clements View author publications You can also search for this author inPubMed Google Scholar * Peter J.

McKinnon View author publications You can also search for this author inPubMed Google Scholar * Keith W. Caldecott View author publications You can also search for this author inPubMed 

Google Scholar * Stephen C. West View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Stephen C. West. ETHICS

DECLARATIONS COMPETING INTERESTS Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION

SUPPLEMENTARY NOTES This file contains Supplementary Methods and Supplementary Figure Legends. (DOC 74 kb) SUPPLEMENTARY FIGURE 1 Reaction mechanism of DNA ligases. (PDF 114 kb)

SUPPLEMENTARY FIGURE 2 Purification of recombinant human Aprataxin. (PDF 257 kb) SUPPLEMENTARY FIGURE 3 Direct measurement of AMP release from the DNA-adenylate intermediate. (PDF 209 kb)

SUPPLEMENTARY FIGURE 4 Analysis of Aprataxin on the DNA ligase-adenylate complex. (PDF 133 kb) SUPPLEMENTARY FIGURE 5 Phylogenetic tree showing that Aprataxin (APTX) forms a distinct branch

of the HIT superfamily. (PDF 111 kb) SUPPLEMENTARY FIGURE 6 Extracts from Aptx-disrupted DT40 cells exhibit reduced ligation activity with the DNA adenylate substrate. (PDF 355 kb)

SUPPLEMENTARY FIGURE 7 Generation and characterization of Aptx-/- mice. (PDF 210 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Ahel, I., Rass, U.,

El-Khamisy, S. _et al._ The neurodegenerative disease protein aprataxin resolves abortive DNA ligation intermediates. _Nature_ 443, 713–716 (2006). https://doi.org/10.1038/nature05164

Download citation * Received: 07 June 2006 * Accepted: 17 August 2006 * Published: 10 September 2006 * Issue Date: 12 October 2006 * DOI: https://doi.org/10.1038/nature05164 SHARE THIS

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