53BP1 ablation rescues genomic instability in mice expressing 'RING-less' BRCA1.
BRCA1 mutations strongly predispose affected individuals to breast and ovarian cancer, but the mechanism by which BRCA1 acts as a tumor suppressor is not fully understood. Homozygous deletion of exon 2 of the mouse Brca1 gene normally causes embryonic lethality, but we show that exon 2-deleted alleles of Brca1 are expressed as a mutant isoform that lacks the N-terminal RING domain. This "RING-less" BRCA1 protein is stable and efficiently recruited to the sites of DNA damage. Surprisingly, robust RAD51 foci form in cells expressing RING-less BRCA1 in response to DNA damage, but the cells nonetheless display the substantial genomic instability. Genomic instability can be rescued by the deletion of Trp53bp1, which encodes the DNA damage response factor 53BP1, and mice expressing RING-less BRCA1 do not show an increased susceptibility to tumors in the absence of 53BP1. Genomic instability in cells expressing RING-less BRCA1 correlates with the loss of BARD1 and a defect in restart of replication forks after hydroxyurea treatment, suggesting a role of BRCA1-BARD1 in genomic integrity that is independent of RAD51 loading.
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Minxing
M
Li
Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA.|Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, TX, USA.
Francesca
F
Cole
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Dharm S
DS
Patel
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Sarah M
SM
Misenko
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Joonyoung
J
Her
Laboratory of Genome Integrity, Center for Cancer Research, National Cancer Institute National Institutes of Health, Bethesda, MD, USA.
Amy
A
Malhowski
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Ali
0000-0001-7666-8531
A
Alhamza
Biological Mass Spectrometry Facility, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Haiyan
H
Zheng
Institute of Cancer Genetics, Department of Pathology & Cell Biology, Columbia University Medical Center, New York, NY, USA.
Richard
R
Baer
Department of Cancer Biology & Genetics, Ohio State University, Columbus, OH, USA.
Thomas
T
Ludwig
Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA.
Maria
M
Jasin
Laboratory of Genome Integrity, Center for Cancer Research, National Cancer Institute National Institutes of Health, Bethesda, MD, USA.
André
A
Nussenzweig
Department of Genetics, Human Genetics Institute of New Jersey, Rutgers The State University of New Jersey, Piscataway, NJ, USA.
Lourdes
0000-0002-3409-9366
L
Serrano
Department of Molecular Biology and Biochemistry, Rutgers The State University of New Jersey, Piscataway, NJ, USA bunting@cabm.rutgers.edu.
Samuel F
0000-0002-4629-496X
SF
Bunting
D000818:Animals; D019313:BRCA1 Protein; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins; D005091:Exons / Q000235:genetics; D005260:Female; D042822:Genomic Instability; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins; D017384:Sequence Deletion; D025521:Tumor Suppressor Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D000071857:Tumor Suppressor p53-Binding Protein 1 / Q000172:deficiency / Q000235:genetics*; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics
D019313:BRCA1 Protein; C000605931:Brca1 protein, mouse; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C106710:Rad51ap1 protein, mouse; C498302:Trp53bp1 protein, mouse; D025521:Tumor Suppressor Proteins; D000071857:Tumor Suppressor p53-Binding Protein 1; C491163:Bard1 protein, mouse; D044767:Ubiquitin-Protein Ligases
DNA repair; RAD51; cancer; genomic integrity; mouse models
J Clin Invest. 2016 Aug 1;126(8):2903-18
27454287
Nature. 2016 Jul 20;535(7612):382-7
27443740
Cold Spring Harb Perspect Biol. 2015 Apr 01;7(4):a016600
25833843
Methods Mol Biol. 2013;927:299-307
22992924
Nat Genet. 1996 Dec;14(4):430-40
8944023
Proc Natl Acad Sci U S A. 2008 May 13;105(19):7040-5
18443292
Nat Struct Mol Biol. 2010 Jun;17(6):688-95
20453858
Cancer Cell. 2011 Dec 13;20(6):797-809
22172724
Cell. 2010 Apr 16;141(2):243-54
20362325
Trends Genet. 2001 Oct;17(10):S18-22
11585672
Proc Natl Acad Sci U S A. 2001 Oct 9;98(21):12078-83
11593018
Nat Cell Biol. 2002 Dec;4(12):993-7
12447390
Proc Natl Acad Sci U S A. 2003 May 13;100(10):5646-51
12732733
Nat Genet. 2001 Jul;28(3):266-71
11431698
J Vis Exp. 2014 Aug 19;(90):
25177909
Mol Cell Biol. 2003 Apr;23(7):2556-63
12640136
Mol Cell. 1999 Mar;3(3):389-95
10198641
Proc Natl Acad Sci U S A. 2008 Dec 30;105(52):20876-81
19088202
J Cell Biol. 2004 May 24;165(4):459-64
15159415
Mol Endocrinol. 2007 Mar;21(3):651-63
17185394
J Clin Invest. 2016 Aug 1;126(8):3145-57
27454289
Cell Rep. 2014 Aug 21;8(4):999-1005
25131202
Mol Cell. 2009 Aug 28;35(4):534-41
19716796
Nature. 2011 Sep 07;477(7363):179-84
21901007
Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7155-60
19369211
Nat Cell Biol. 2012 Mar 04;14(4):424-30
22388890
Curr Biol. 2004 Dec 14;14(23):2135-42
15589157
Annu Rev Genet. 2015;49:395-412
26631513
Cancer Cell. 2012 Jul 10;22(1):106-16
22789542
EMBO Rep. 2016 Nov;17(11):1532-1541
27670884
Mol Cell. 1999 Oct;4(4):511-8
10549283
Cell. 2013 Jan 31;152(3):620-32
23352430
Nucleic Acids Res. 1997 Mar 15;25(6):1317-8
9092650
Oncogene. 2006 Sep 25;25(43):5885-97
16998503
EMBO J. 2016 Jul 15;35(14):1488-503
27225932
Genes Dev. 1997 May 15;11(10):1226-41
9171368
Science. 2011 Oct 28;334(6055):525-8
22034435
Nat Rev Cancer. 2011 Dec 23;12(1):68-78
22193408
J Proteome Res. 2004 Nov-Dec;3(6):1234-42
15595733
J Biol Chem. 2001 May 4;276(18):14537-40
11278247
Mol Cell. 2012 Apr 27;46(2):125-35
22445484