@article{9140a11380ff4c348f525f8458ef97ae,
title = "Crystal Structure of the First Eubacterial Mre11 Nuclease Reveals Novel Features that May Discriminate Substrates During DNA Repair",
abstract = "Mre11 nuclease plays a central role in the repair of cytotoxic and mutagenic DNA double-strand breaks. As X-ray structural information has been available only for the Pyrococcus furiosus enzyme (PfMre11), the conserved and variable features of this nuclease across the domains of life have not been experimentally defined. Our crystal structure and biochemical studies demonstrate that TM1635 from Thermotoga maritima, originally annotated as a putative nuclease, is an Mre11 endo/exonuclease (TmMre11) and the first such structure from eubacteria. TmMre11 and PfMre11 display similar overall structures, despite sequence identity in the twilight zone of only ∼20%. However, they differ substantially in their DNA-specificity domains and in their dimeric organization. Residues in the nuclease domain are highly conserved, but those in the DNA-specificity domain are not. The structural differences likely affect how Mre11 from different organisms recognize and interact with single-stranded DNA, double-stranded DNA and DNA hairpin structures during DNA repair. The TmMre11 nuclease active site has no bound metal ions, but is conserved in sequence and structure with the exception of a histidine that is important in PfMre11 nuclease activity. Nevertheless, biochemical characterization confirms that TmMre11 possesses both endonuclease and exonuclease activities on single-stranded and double-stranded DNA substrates, respectively.",
keywords = "DNA repair, Mre11, crystal structure, nuclease, structural genomics",
author = "Debanu Das and Davide Moiani and Axelrod, {Herbert L.} and Miller, {Mitchell D.} and Daniel McMullan and Jin, {Kevin K.} and Polat Abdubek and Tamara Astakhova and Prasad Burra and Dennis Carlton and Chiu, {Hsiu Ju} and Thomas Clayton and Deller, {Marc C.} and Lian Duan and Dustin Ernst and Julie Feuerhelm and Grant, {Joanna C.} and Anna Grzechnik and Grzechnik, {Slawomir K.} and Han, {Gye Won} and Lukasz Jaroszewski and Klock, {Heath E.} and Knuth, {Mark W.} and Piotr Kozbial and Krishna, {S. Sri} and Abhinav Kumar and David Marciano and Morse, {Andrew T.} and Edward Nigoghossian and Linda Okach and Jessica Paulsen and Ron Reyes and Rife, {Christopher L.} and Natasha Sefcovic and Tien, {Henry J.} and Trame, {Christine B.} and {van den Bedem}, Henry and Dana Weekes and Qingping Xu and Hodgson, {Keith O.} and John Wooley and Elsliger, {Marc Andr{\'e}} and Deacon, {Ashley M.} and Adam Godzik and Lesley, {Scott A.} and Tainer, {John A.} and Wilson, {Ian A.}",
note = "Funding Information: The project was sponsored by the National Institute of General Medical Sciences Protein Structure Initiative (P50 GM62411, U54 GM074898). Portions of this research were performed at the Stanford Synchrotron Radiation Lightsource (SSRL). The SSRL is a national user facility at the SLAC National Accelerator Laboratory operated by Stanford University on behalf of the United States Department of Energy, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research, and by the National Institutes of Health (National Center for Research Resources, Biomedical Technology Program, and the National Institute of General Medical Sciences). Mre11 research in the Tainer group is funded by CA117638 and CA092584. We are grateful to Chiharu Hitomi and Dr Julie Tubbs in the Tainer group for assistance with site-directed mutagenesis and biochemical assays. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the National Institutes of Health. Genomic DNA from Thermotoga maritima MSB8 (DSM3109) (ATCC #43589D-5) was obtained from the American Type Culture Collection (ATCC). ",
year = "2010",
month = apr,
day = "2",
doi = "10.1016/j.jmb.2010.01.049",
language = "English (US)",
volume = "397",
pages = "647--663",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "3",
}