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PMID: 17349953 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Rad50 adenylate kinase activity regulates DNA tethering by Mre11/Rad50 complexes.

Molecular cell ·Vol. 25 ·No. 5 ·2007-03-09 ·Pages 647-61

Bhaskara V, Dupré A, Lengsfeld B, Hopkins BB, Chan A, Lee JH, Zhang X, Gautier J, Zakian V, Paull TT

Abstract

Mre11 and Rad50 are the catalytic components of a highly conserved DNA repair complex that functions in many aspects of DNA metabolism involving double-strand breaks. The ATPase domains in Rad50 are related to the ABC transporter family of ATPases, previously shown to share structural similarities with adenylate kinases. Here we demonstrate that Mre11/Rad50 complexes from three organisms catalyze the reversible adenylate kinase reaction in vitro. Mutation of the conserved signature motif reduces the adenylate kinase activity of Rad50 but does not reduce ATP hydrolysis. This mutant resembles a rad50 null strain with respect to meiosis and telomere maintenance in S. cerevisiae, correlating adenylate kinase activity with in vivo functions. An adenylate kinase inhibitor blocks Mre11/Rad50-dependent DNA tethering in vitro and in cell-free extracts, indicating that adenylate kinase activity by Mre11/Rad50 promotes DNA-DNA associations. We propose a model for Rad50 that incorporates both ATPase and adenylate kinase reactions as critical activities that regulate Rad50 functions.

MeSH Terms
Acid Anhydride Hydrolases Adenine/metabolism Adenosine Triphosphate/metabolism Adenylate Kinase/antagonists & inhibitors,metabolism Amino Acid Motifs Animals Archaeal Proteins/metabolism Catalysis/drug effects DNA/metabolism DNA Repair Enzymes/metabolism DNA-Binding Proteins/metabolism Dinucleoside Phosphates/metabolism Endodeoxyribonucleases/metabolism Enzyme Inhibitors/pharmacology Exodeoxyribonucleases/metabolism Humans Hydrolysis/drug effects MRE11 Homologue Protein Mutant Proteins/metabolism Mutation/genetics Pyrococcus furiosus/drug effects,enzymology Saccharomyces cerevisiae/drug effects,enzymology Saccharomyces cerevisiae Proteins/metabolism Xenopus
Chemicals
Archaeal Proteins DNA-Binding Proteins Dinucleoside Phosphates Enzyme Inhibitors MRE11 protein, human Mutant Proteins RAD50 protein, S cerevisiae Saccharomyces cerevisiae Proteins P(1),P(5)-di(adenosine-5'-)pentaphosphate Adenosine Triphosphate DNA Adenylate Kinase Endodeoxyribonucleases Exodeoxyribonucleases MRE11 Homologue Protein MRE11 protein, S cerevisiae Mre11 protein, archaeal Rad50 protein, archaeal Acid Anhydride Hydrolases Rad50 protein, human DNA Repair Enzymes Adenine
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Bhaskara Venugopal
Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX 78712, USA.
Dupré Aude
Lengsfeld Bettina
Hopkins Ben B
Chan Angela
Lee Ji-Hoon
Zhang Xiaoming
Gautier Jean
Zakian Virginia
Paull Tanya T
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-2765
DOI
10.1016/j.molcel.2007.01.028
Published
2007-03-09
Pages
647-61
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC3050042
Subset
IM
Grants
NCI NIH HHS · R01 CA092245-10 · United States
NCI NIH HHS · R01 CA094008-09 · United States
NCI NIH HHS · R01 CA092245 · United States
NCI NIH HHS · R01 CA092245-06 · United States
NCI NIH HHS · R01 CA 094008 · United States
NCI NIH HHS · R01 CA 92245 · United States
NCI NIH HHS · R01 CA094008 · United States
Corrections
CommentIn
PMID: 17386254
External Links
PubMed source DOI: 10.1016/j.molcel.2007.01.028 PMC Full Text View journal details
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