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Structural and functional insights into DNA-end processing by the archaeal HerA helicase-NurA nuclease complex

Published version
Peer-reviewed

Type

Article

Change log

Authors

Blackwood, JK 
Rzechorzek, NJ 
Abrams, AS 
Maman, JD 

Abstract

Helicase-nuclease systems dedicated to DNA end resection in preparation for homologous recombination (HR) are present in all kingdoms of life. In thermophilic archaea, the HerA helicase and NurA nuclease cooperate with the highly conserved Mre11 and Rad50 proteins during HR-dependent DNA repair. Here we show that HerA and NurA must interact in a complex with specific subunit stoichiometry to process DNA ends efficiently. We determine crystallographically that NurA folds in a toroidal dimer of intertwined RNaseH-like domains. The central channel of the NurA dimer is too narrow for double-stranded DNA but appears well suited to accommodate one or two strands of an unwound duplex. We map a critical interface of the complex to an exposed hydrophobic epitope of NurA abutting the active site. Based upon the presented evidence, we propose alternative mechanisms of DNA end processing by the HerA-NurA complex.

Description

Keywords

Adenosine Triphosphate, Amino Acid Sequence, Archaeal Proteins, Conserved Sequence, Crystallography, X-Ray, DNA, DNA Helicases, Deoxyribonucleases, Dimerization, Models, Molecular, Molecular Sequence Data, Protein Folding, Protein Structure, Tertiary, Ribonuclease H, Sulfolobus solfataricus

Journal Title

Nucleic Acids Research

Conference Name

Journal ISSN

0305-1048
1362-4962

Volume Title

40

Publisher

Oxford University Press
Sponsorship
Medical Research Council (G0701443)
Wellcome Trust (084279/Z/07/A)
Wellcome Trust (084279/Z/07/Z)
Research in the N.P.R. laboratory is funded by the Medical Research Council [Career Development Award (G0701443)]. Research in L.P. laboratory is funded by a Wellcome Trust Senior Fellowship Award in Basic Biomedical Sciences (08279/Z/07/Z). Funding for open access charge: the Medical Research (G0701443 to N.P.R.) and the Wellcome Trust (08279/Z/07/Z to L.P.) and the Department of Biochemistry, University of Cambridge.