title: A winged helix domain in human MUS81 binds DNA and modulates the endonuclease activity of MUS81 complexes.
abstract: The MUS81-EME1 endonuclease maintains metazoan genomic integrity by cleaving branched DNA structures that arise during the resolution of recombination intermediates. In humans, MUS81 also forms a poorly characterized complex with EME2. Here, we identify and determine the structure of a winged helix (WH) domain from human MUS81, which binds DNA. WH domain mutations greatly reduce binding of the isolated domain to DNA and impact on incision activity of MUS81-EME1/EME2 complexes. Deletion of the WH domain reduces the endonuclease activity of both MUS81-EME1 and MUS81-EME2 complexes, and incisions made by MUS81-EME2 are made closer to the junction on substrates containing a downstream duplex, such as fork structures and nicked Holliday junctions. WH domain mutation or deletion in Schizosaccharomyces pombe phenocopies the DNA-damage sensitivity of strains deleted for mus81. Our results indicate an important role for the WH domain in both yeast and human MUS81 complexes.
authors: Structural Biology Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3LY, UK, Genome Damage and Stability Centre, University of Sussex, Brighton, BN1 9RQ, UK and Institute of Structural and Molecular Biology, University College London and Birkbeck College, Malet Street, London WC1E 7HX, UK.
authors: Andrew J
authors: AJ
authors: Fadden
authors: Stephanie
authors: S
authors: Schalbetter
authors: Maureen
authors: M
authors: Bowles
authors: Richard
authors: R
authors: Harris
authors: John
authors: J
authors: Lally
authors: Antony M
authors: AM
authors: Carr
authors: Neil Q
authors: NQ
authors: McDonald
mesh_terms: D000595:Amino Acid Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004720:Endonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary
chemical_list: D004268:DNA-Binding Proteins; D004247:DNA; D004706:Endodeoxyribonucleases; D004720:Endonucleases; C498772:MUS81 protein, human
references: Mol Biol Cell. 2004 Feb;15(2):552-62
references: 14617801
references: Nat Protoc. 2009;4(3):363-71
references: 19247286
references: Cell. 2001 Nov 16;107(4):537-48
references: 11719193
references: J Biomol NMR. 1999 Mar;13(3):289-302
references: 10212987
references: Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
references: 9757107
references: Structure. 2005 Aug;13(8):1183-92
references: 16084390
references: EMBO J. 2002 Apr 15;21(8):2045-53
references: 11953324
references: J Biomol NMR. 1999 Jan;13(1):51-9
references: 10905826
references: Mol Cell. 2007 Feb 9;25(3):331-43
references: 17289582
references: Curr Opin Struct Biol. 2000 Feb;10(1):110-6
references: 10679470
references: Biometals. 2005 Aug;18(4):413-28
references: 16158234
references: J Magn Reson. 1998 Apr;131(2):373-8
references: 9571116
references: DNA Repair (Amst). 2007 Jul 1;6(7):1004-17
references: 17409028
references: EMBO J. 2005 Mar 9;24(5):895-905
references: 15719018
references: Nucleic Acids Res. 2010 Nov;38(21):7611-25
references: 20660481
references: Cell. 2009 Jul 10;138(1):63-77
references: 19596235
references: Gene. 2008 Jan 15;407(1-2):63-74
references: 18054176
references: Nucleic Acids Res. 2009 Apr;37(6):2026-36
references: 19211663
references: FEBS J. 2012 Jul;279(13):2412-30
references: 22551069
references: EMBO J. 2002 Oct 15;21(20):5577-85
references: 12374758
references: Mol Cell. 2001 Nov;8(5):1117-27
references: 11741546
references: J Biomol NMR. 2001 Jul;20(3):223-31
references: 11519746
references: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3757-62
references: 18310322
references: Nature. 2008 Nov 20;456(7220):357-61
references: 19020614
references: Genes Dev. 2008 Apr 15;22(8):1093-106
references: 18413719
references: Nature. 1998 Jul 30;394(6692):502-6
references: 9697776
references: Cell. 2009 Jul 10;138(1):78-89
references: 19596236
references: J Mol Biol. 1999 Sep 24;292(3):653-67
references: 10497029
references: DNA Repair (Amst). 2003 Dec 9;2(12):1435-47
references: 14642571
references: J Biomol NMR. 1995 Nov;6(3):277-93
references: 8520220
references: Structure. 2005 Dec;13(12):1849-58
references: 16338413
references: Cancer Res. 2005 Apr 1;65(7):2526-31
references: 15805243
references: Mol Cell. 2009 Jul 10;35(1):116-27
references: 19595721
references: J Biomol NMR. 1995 Jan;5(1):67-81
references: 7881273
references: J Biol Chem. 2003 Jul 4;278(27):25172-8
references: 12721304
references: Genes Dev. 1997 Dec 15;11(24):3375-86
references: 9407030
references: Annu Rev Biochem. 2008;77:259-87
references: 18518821