The de-ubiquitylating enzymes USP26 and USP37 regulate homologous recombination by counteracting RAP80

TY - JOUR

T1 - The de-ubiquitylating enzymes USP26 and USP37 regulate homologous recombination by counteracting RAP80

AU - Typas, Dimitris

AU - Luijsterburg, Martijn S

AU - Wiegant, Wouter W

AU - Diakatou, Michaela

AU - Helfricht, Angela

AU - Thijssen, Peter E

AU - van den Broek, Bram

AU - van de Broek, Bram

AU - Mullenders, Leon H

AU - van Attikum, Haico

N1 - © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2015/8/18

Y1 - 2015/8/18

N2 - The faithful repair of DNA double-strand breaks (DSBs) is essential to safeguard genome stability. DSBs elicit a signaling cascade involving the E3 ubiquitin ligases RNF8/RNF168 and the ubiquitin-dependent assembly of the BRCA1-Abraxas-RAP80-MERIT40 complex. The association of BRCA1 with ubiquitin conjugates through RAP80 is known to be inhibitory to DSB repair by homologous recombination (HR). However, the precise regulation of this mechanism remains poorly understood. Through genetic screens we identified USP26 and USP37 as key de-ubiquitylating enzymes (DUBs) that limit the repressive impact of RNF8/RNF168 on HR. Both DUBs are recruited to DSBs where they actively remove RNF168-induced ubiquitin conjugates. Depletion of USP26 or USP37 disrupts the execution of HR and this effect is alleviated by the simultaneous depletion of RAP80. We demonstrate that USP26 and USP37 prevent excessive spreading of RAP80-BRCA1 from DSBs. On the other hand, we also found that USP26 and USP37 promote the efficient association of BRCA1 with PALB2. This suggests that these DUBs limit the ubiquitin-dependent sequestration of BRCA1 via the BRCA1-Abraxas-RAP80-MERIT40 complex, while promoting complex formation and cooperation of BRCA1 with PALB2-BRCA2-RAD51 during HR. These findings reveal a novel ubiquitin-dependent mechanism that regulates distinct BRCA1-containing complexes for efficient repair of DSBs by HR.

AB - The faithful repair of DNA double-strand breaks (DSBs) is essential to safeguard genome stability. DSBs elicit a signaling cascade involving the E3 ubiquitin ligases RNF8/RNF168 and the ubiquitin-dependent assembly of the BRCA1-Abraxas-RAP80-MERIT40 complex. The association of BRCA1 with ubiquitin conjugates through RAP80 is known to be inhibitory to DSB repair by homologous recombination (HR). However, the precise regulation of this mechanism remains poorly understood. Through genetic screens we identified USP26 and USP37 as key de-ubiquitylating enzymes (DUBs) that limit the repressive impact of RNF8/RNF168 on HR. Both DUBs are recruited to DSBs where they actively remove RNF168-induced ubiquitin conjugates. Depletion of USP26 or USP37 disrupts the execution of HR and this effect is alleviated by the simultaneous depletion of RAP80. We demonstrate that USP26 and USP37 prevent excessive spreading of RAP80-BRCA1 from DSBs. On the other hand, we also found that USP26 and USP37 promote the efficient association of BRCA1 with PALB2. This suggests that these DUBs limit the ubiquitin-dependent sequestration of BRCA1 via the BRCA1-Abraxas-RAP80-MERIT40 complex, while promoting complex formation and cooperation of BRCA1 with PALB2-BRCA2-RAD51 during HR. These findings reveal a novel ubiquitin-dependent mechanism that regulates distinct BRCA1-containing complexes for efficient repair of DSBs by HR.

KW - BRCA1 Protein

KW - Carrier Proteins

KW - Cell Line

KW - Cysteine Endopeptidases

KW - DNA Breaks, Double-Stranded

KW - Endopeptidases

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - Nuclear Proteins

KW - Rad51 Recombinase

KW - Recombinational DNA Repair

KW - Ubiquitin

KW - Ubiquitin-Protein Ligases

KW - Ubiquitination

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1093/nar/gkv613

DO - 10.1093/nar/gkv613

M3 - Journal article

C2 - 26101254

SN - 0305-1048

VL - 43

SP - 6919

EP - 6933

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 14

ER -