Phosphorylated CtIP Functions as a Co-factor of the MRE11-RAD50-NBS1 Endonuclease in DNA End Resection

• DOI: 10.1016/j.molcel.2016.10.017
• Publication Date: 2016-11-23T21:16:41Z
• Authors: Roopesh Anand; Lepakshi Ranjha; Elda Cannavo; Petr Cejka
• Keywords: 0301 basic medicine; MRE11 Homologue Protein; 0303 health sciences; DNA End-Joining Repair; Endodeoxyribonucleases; 10061 Institute of Molecular Cancer Research; DNA Helicases; Nuclear Proteins; 610 Medicine & health; Cell Cycle Proteins; Recombinant Proteins; Acid Anhydride Hydrolases; 3. Good health; 1307 Cell Biology; DNA-Binding Proteins; 03 medical and health sciences; DNA Repair Enzymes; Multiprotein Complexes; 1312 Molecular Biology; 570 Life sciences; biology; Humans; DNA Breaks, Double-Stranded; Phosphorylation; Carrier Proteins; Homologous Recombination

To repair a DNA double-strand break (DSB) by homologous recombination (HR), the 5'-terminated strand of the DSB must be resected. The human MRE11-RAD50-NBS1 (MRN) and CtIP proteins were implicated in the initiation of DNA end resection, but the underlying mechanism remained undefined. Here, we show that CtIP is a co-factor of the MRE11 endonuclease activity within the MRN complex. This function is absolutely dependent on CtIP phosphorylation that includes the key cyclin-dependent kinase target motif at Thr-847. Unlike in yeast, where the Xrs2/NBS1 subunit is dispensable in vitro, NBS1 is absolutely required in the human system. The MRE11 endonuclease in conjunction with RAD50, NBS1, and phosphorylated CtIP preferentially cleaves 5'-terminated DNA strands near DSBs. Our results define the initial step of HR that is particularly relevant for the processing of DSBs bearing protein blocks.