RAD51D splice variants and cancer-associated mutations reveal XRCC2 interaction to be critical for homologous recombination.
Double-strand break repair
Homologous recombination
RAD51 paralogs
RAD51D
Walker A motif
XRCC2
Journal
DNA repair
ISSN: 1568-7856
Titre abrégé: DNA Repair (Amst)
Pays: Netherlands
ID NLM: 101139138
Informations de publication
Date de publication:
04 2019
04 2019
Historique:
received:
27
08
2018
revised:
15
02
2019
accepted:
15
02
2019
pubmed:
6
3
2019
medline:
17
7
2019
entrez:
6
3
2019
Statut:
ppublish
Résumé
The proficiency of cancer cells to repair DNA double-strand breaks (DSBs) by homologous recombination (HR) is a key determinant in predicting response to targeted therapies such as PARP inhibitors. The RAD51 paralogs work as multimeric complexes and act downstream of BRCA1 to facilitate HR. Numerous epidemiological studies have linked RAD51 paralog mutations with hereditary cancer predisposition. Despite their substantial links to cancer, RAD51 paralog HR function has remained elusive. Here we identify isoform 1 as the functional isoform of RAD51D, whereas isoform 4 which has a large N-terminal deletion (including the Walker A motif), and isoform 6 which includes an alternate exon in the N-terminus, are non-functional. To determine the importance of this N-terminal region, we investigated the impact of cancer-associated mutations and SNPs in this variable RAD51D N-terminal region using yeast-2-hybrid and yeast-3-hybrid assays to screen for altered protein-protein interactions. We identified two cancer-associated mutations close to or within the Walker A motif (G96C and G107 V, respectively) that independently disrupt RAD51D interaction with XRCC2. We validated our yeast interaction data in human U2OS cells by co-immunoprecipitation and determined the impact of these mutations on HR-proficiency using a sister chromatid recombination reporter assay in a RAD51D knock-out cell line. Our investigation reveals that the interaction of RAD51D with XRCC2 is required for DSB repair. By characterizing the impact of cancer-associated mutations on RAD51D interactions, we aim to develop predictive models for therapeutic sensitivity and resistance in patients who harbor similar mutations in RAD51D.
Identifiants
pubmed: 30836272
pii: S1568-7864(18)30193-9
doi: 10.1016/j.dnarep.2019.02.008
pmc: PMC6508892
mid: NIHMS1523067
pii:
doi:
Substances chimiques
DNA-Binding Proteins
0
Protein Isoforms
0
RAD51D protein, human
0
XRCC2 protein, human
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
99-107Subventions
Organisme : NIGMS NIH HHS
ID : F32 GM110978
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA185660
Pays : United States
Organisme : NIEHS NIH HHS
ID : R01 ES024872
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA047904
Pays : United States
Organisme : NIGMS NIH HHS
ID : K99 GM088413
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA008748
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM118175
Pays : United States
Organisme : NIGMS NIH HHS
ID : R00 GM088413
Pays : United States
Informations de copyright
Copyright © 2019 Elsevier B.V. All rights reserved.
Références
Mol Cell. 1999 Oct;4(4):511-8
pubmed: 10549283
Mol Cell Biol. 1999 Dec;19(12):8686-93
pubmed: 10567591
J Biol Chem. 2000 Jun 2;275(22):16443-9
pubmed: 10749867
EMBO J. 2000 Dec 15;19(24):6675-85
pubmed: 11118202
Nucleic Acids Res. 2001 Jan 1;29(1):308-11
pubmed: 11125122
J Biol Chem. 2001 Jun 22;276(25):22148-53
pubmed: 11301337
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8644-9
pubmed: 11447276
Genes Dev. 2001 Dec 15;15(24):3296-307
pubmed: 11751635
J Biol Chem. 2002 Apr 19;277(16):14315-20
pubmed: 11834724
Nucleic Acids Res. 2004 Jan 02;32(1):169-78
pubmed: 14704354
Cell. 2004 Apr 30;117(3):337-47
pubmed: 15109494
DNA Repair (Amst). 2005 Feb 3;4(2):149-61
pubmed: 15590323
Nature. 2005 Apr 14;434(7035):913-7
pubmed: 15829966
Nature. 2005 Apr 14;434(7035):917-21
pubmed: 15829967
Mutagenesis. 2005 Nov;20(6):433-40
pubmed: 16236763
EMBO J. 2006 Jun 7;25(11):2564-74
pubmed: 16710300
Nucleic Acids Res. 2006 May 22;34(9):2833-43
pubmed: 16717288
Cancer Res. 2009 Feb 1;69(3):863-72
pubmed: 19155299
BMC Mol Biol. 2009 Mar 27;10:27
pubmed: 19327148
Nat Methods. 2010 Apr;7(4):248-9
pubmed: 20354512
Lancet. 2010 Jul 24;376(9737):235-44
pubmed: 20609467
Lancet. 2010 Jul 24;376(9737):245-51
pubmed: 20609468
Sci Signal. 2011 Jun 28;4(179):rs5
pubmed: 21712546
Semin Cell Dev Biol. 2011 Oct;22(8):898-905
pubmed: 21821141
Lancet Oncol. 2011 Sep;12(9):852-61
pubmed: 21862407
J Biol Chem. 2011 Dec 2;286(48):41758-66
pubmed: 21965664
Nucleic Acids Res. 2012 Jan;40(Database issue):D261-70
pubmed: 22135298
Nat Genet. 2012 Oct;44(10):1104-10
pubmed: 22941188
Mol Cell Biol. 2013 Jan;33(2):387-95
pubmed: 23149936
PLoS One. 2013;8(1):e54772
pubmed: 23372765
Mol Cell Biol. 2013 May;33(9):1830-44
pubmed: 23438602
Cold Spring Harb Perspect Biol. 2013 Nov 01;5(11):a012740
pubmed: 24097900
Oncogene. 2014 Oct 2;33(40):4803-12
pubmed: 24141787
PLoS One. 2013 Dec 05;8(12):e81371
pubmed: 24339919
Nat Rev Mol Cell Biol. 2014 Jun;15(6):369-83
pubmed: 24824069
Nature. 2015 Jan 29;517(7536):576-82
pubmed: 25631445
Springerplus. 2015 Feb 24;4:92
pubmed: 25918678
Nature. 2015 Aug 6;524(7563):47-53
pubmed: 26168399
J Clin Oncol. 2015 Sep 10;33(26):2901-7
pubmed: 26261251
Am J Cancer Res. 2015 Jun 15;5(7):2113-26
pubmed: 26328243
Nucleic Acids Res. 2015 Nov 16;43(20):9835-55
pubmed: 26354865
Cancer Biomark. 2015;15(6):775-81
pubmed: 26406419
Genetics. 2016 May;203(1):133-45
pubmed: 26936927
J Clin Oncol. 2016 May 1;34(13):1460-8
pubmed: 26976419
PLoS One. 2016 May 05;11(5):e0153788
pubmed: 27149063
DNA Repair (Amst). 2016 Jun;42:82-93
pubmed: 27161866
PLoS One. 2016 May 19;11(5):e0155476
pubmed: 27195665
Biochem Cell Biol. 2016 Oct;94(5):407-418
pubmed: 27224545
Nature. 2016 May 25;534(7605):55-62
pubmed: 27251275
Nucleic Acids Res. 2016 Sep 30;44(17):8199-215
pubmed: 27298254
Lancet Oncol. 2017 Jan;18(1):75-87
pubmed: 27908594
Nucleic Acids Res. 2017 Feb 28;45(4):1835-1847
pubmed: 27924006
Breast Cancer Res Treat. 2017 Jun;163(2):383-390
pubmed: 28281021
JAMA Oncol. 2017 Sep 1;3(9):1190-1196
pubmed: 28418444
Ann Surg Oncol. 2017 Oct;24(10):3060-3066
pubmed: 28766213
Genomics. 1998 Apr 1;49(1):103-11
pubmed: 9570954
Genes Dev. 1998 Dec 15;12(24):3831-42
pubmed: 9869637