Coordination of structure-specific nucleases by human SLX4/BTBD12 is required for DNA repair.
Budding yeast Slx4 interacts with the structure-specific endonuclease Slx1 to ensure completion of ribosomal DNA replication. Slx4 also interacts with the Rad1-Rad10 endonuclease to control cleavage of 3' flaps during repair of double-strand breaks (DSBs). Here we describe the identification of human SLX4, a scaffold for DNA repair nucleases XPF-ERCC1, MUS81-EME1, and SLX1. SLX4 immunoprecipitates show SLX1-dependent nuclease activity toward Holliday junctions and MUS81-dependent activity toward other branched DNA structures. Furthermore, SLX4 enhances the nuclease activity of SLX1, MUS81, and XPF. Consistent with a role in processing recombination intermediates, cells depleted of SLX4 are hypersensitive to genotoxins that cause DSBs and show defects in the resolution of interstrand crosslink-induced DSBs. Depletion of SLX4 causes a decrease in DSB-induced homologous recombination. These data show that SLX4 is a regulator of structure-specific nucleases and that SLX4 and SLX1 are important regulators of genome stability in human cells.
Muñoz
Ivan M
IM
MRC Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Hain
Karolina
K
Déclais
Anne-Cécile
AC
Gardiner
Mary
M
Toh
Geraldine W
GW
Sanchez-Pulido
Luis
L
Heuckmann
Johannes M
JM
Toth
Rachel
R
Macartney
Thomas
T
Eppink
Berina
B
Kanaar
Roland
R
Ponting
Chris P
CP
Lilley
David M J
DM
Rouse
John
J
D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics; D045522:Recombinases / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques
D004268:DNA-Binding Proteins; D034741:RNA, Small Interfering; D045522:Recombinases; C095888:xeroderma pigmentosum group F protein; D049452:Green Fluorescent Proteins; C071447:ERCC1 protein, human; D004720:Endonucleases; C498772:MUS81 protein, human; C542319:SLX4 protein, human