BRIT1/MCPH1 Is Essential for Mitotic and Meiotic Recombination DNA Repair and Maintaining Genomic Stability in Mice

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BRIT1 protein (also known as MCPH1) contains 3 BRCT domains which are conserved in BRCA1, BRCA2, and other important molecules involved in DNA damage signaling, DNA repair, and tumor suppression. BRIT1 mutations or aberrant expression are found in primary microcephaly patients as well as in cancer patients. Recent in vitro studies suggest that BRIT1/MCPH1 functions as a novel key regulator in the DNA damage response pathways. To investigate its physiological role and dissect the underlying mechanisms, we generated BRIT1^−/− mice and identified its essential roles in mitotic and meiotic recombination DNA repair and in maintaining genomic stability. Both BRIT1^−/− mice and mouse embryonic fibroblasts (MEFs) were hypersensitive to γ-irradiation. BRIT1^−/− MEFs and T lymphocytes exhibited severe chromatid breaks and reduced RAD51 foci formation after irradiation. Notably, BRIT1^−/− mice were infertile and meiotic homologous recombination was impaired. BRIT1-deficient spermatocytes exhibited a failure of chromosomal synapsis, and meiosis was arrested at late zygotene of prophase I accompanied by apoptosis. In mutant spermatocytes, DNA double-strand breaks (DSBs) were formed, but localization of RAD51 or BRCA2 to meiotic chromosomes was severely impaired. In addition, we found that BRIT1 could bind to RAD51/BRCA2 complexes and that, in the absence of BRIT1, recruitment of RAD51 and BRCA2 to chromatin was reduced while their protein levels were not altered, indicating that BRIT1 is involved in mediating recruitment of RAD51/BRCA2 to the damage site. Collectively, our BRIT1-null mouse model demonstrates that BRIT1 is essential for maintaining genomic stability in vivo to protect the hosts from both programmed and irradiation-induced DNA damages, and its depletion causes a failure in both mitotic and meiotic recombination DNA repair via impairing RAD51/BRCA2's function and as a result leads to infertility and genomic instability in mice.

Vyšlo v časopise: BRIT1/MCPH1 Is Essential for Mitotic and Meiotic Recombination DNA Repair and Maintaining Genomic Stability in Mice. PLoS Genet 6(1): e32767. doi:10.1371/journal.pgen.1000826
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1000826

Souhrn

Zdroje

1. PastinkA

EekenJC

LohmanPH

2001 Genomic integrity and the repair of double-strand DNA breaks. Mutat Res 480–481 37 50

2. FrancoS

AltFW

ManisJP

2006 Pathways that suppress programmed DNA breaks from progressing to chromosomal breaks and translocations. DNA Repair 5 1030 1041

3. PanditaTK

HittelmanWN

1992 The contribution of DNA and chromosome repair deficiencies to the radiosensitivity of ataxia-telangiectasia. Radiat Res 131 214 223

4. KuzminovA

1995 Collapse and repair of replication forks in Escherichia coli. Mol Microbiol 16 373 384

5. RichardsonC

HorikoshiN

PanditaTK

2004 The role of the DNA double-strand break response network in meiosis. DNA Repair 3 1149 1164

6. RooneyS

ChaudhuriJ

Alt.FW

2004 The role of the non-homologous end-joining pathway in lymphocyte development. Immunol Rev 200 115 131

7. ZhouBB

ElledgeSJ

2000 The DNA damage response: putting checkpoints in perspective. Nature 408 433 439

8. RouseJ

JacksonSP

2002 Interfaces between the detection, signaling, and repair of DNA damage. Science 297 547 551

9. StuckiM

ClappertonJA

MohammadD

YaffeMB

SmerdonSJ

2005 MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell 123 1213 1226

10. PaullTT

RogakouEP

YamazakiV

KirchgessnerCU

GellertM

2000 A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol 10 886 895

11. ChapmanJR

JacksonSP

2008 Phospho-dependent interactions between NBS1 and MDC1 mediate chromatin retention of the MRN complex at sites of DNA damage. EMBO Rep 9 795 801

12. SchultzLB

ChehabNH

MalikzayA

HalazonetisTD

2000 p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J Cell Biol 151 1381 1390

13. RappoldI

IwabuchiK

DateT

ChenJ

2001 Tumor suppressor p53 binding protein 1 (53BP1) is involved in DNA damage-signaling pathways. J Cell Biol 153 613 620

14. SungP

KrejciL

Van KomenS

SehornMG

2003 Rad51 recombinase and recombination mediators. J Biol Chem 278 42729 42732

15. ScullyR

ChenJ

OchsRL

KeeganK

HoekstraM

1997 Dynamic changes of BRCA1 subnuclear location and phosphorylation state are initiated by DNA damage. Cell 90 425 435

16. ThorslundT

WestSC

2007 BRCA2: a universal recombinase regulator. Oncogene 26 7720 7730

17. LinSY

RaiR

LiK

XuZX

ElledgeSJ

2005 BRIT1/MCPH1 is a DNA damage responsive protein that regulates the Brca1-Chk1 pathway, implicating checkpoint dysfunction in microcephaly. Proc Natl Acad Sci USA 102 15105 15109

18. RaiR

DaiH

MultaniAS

LiK

ChinK

2006 BRIT1 regulates early DNA damage response, chromosomal integrity, and cancer. Cancer Cell 10 145 157

19. JacksonAP

EastwoodH

BellSM

AduJ

ToomesC

2002 Identification of microcephalin, a protein implicated in determining the size of the human brain. Am J Hum Genet 71 136 142

20. XuX

LeeJ

SternDF

2004 Microcephalin is a DNA damage response protein involved in regulation of CHK1 and BRCA1. J Biol Chem 279 34091 34094

21. AldertonGK

GalbiatiL

GriffithE

SurinyaKH

NeitzelH

2006 Regulation of mitotic entry by microcephalin and its overlap with ATR signalling. Nat Cell Biol 8 725 733

22. SonodaE

SasakiMS

MorrisonC

Yamaguchi-IwaiY

TakataM

1999 Sister chromatid exchanges are mediated by homologous recombination in vertebrate cells. Mol Cell Biol 19 5166 5169

23. González-BarreraS

Cortés-LedesmaF

WellingerRE

AguileraA

2003 Equal sister chromatid exchange is a major mechanism of double-strand break repair in yeast. Mol Cell 11 1661 1671

24. WoodJL

LiangY

LiK

ChenJ

2008 Microcephalin/MCPH1 associates with the Condensin II complex to function in homologous recombination repair. J Biol Chem 283 29586 29592

25. PengG

YimEK

DaiH

JacksonAP

BurgtI

2009 BRIT1/MCPH1 links chromatin remodelling to DNA damage response. Nat Cell Biol 11 865 872

26. BellvéAR

CavicchiaJC

MilletteCF

O'BrienDA

BhatnagarYM

1977 Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization. J Cell Biol 74 68 85

27. ZhaoGQ

GarbersDL

2002 Male germ cell specification and differentiation. Dev Cell 2 537 547

28. DobsonMJ

PearlmanRE

KaraiskakisA

SpyropoulosB

MoensPB

1994 Synaptonemal complex proteins: occurrence, epitope mapping and chromosome disjunction. J Cell Sci 107 2749 2760

29. SchalkJA

DietrichAJ

VinkAC

OffenbergHH

van AalderenM

1998 Localization of SCP2 and SCP3 protein molecules within synaptonemal complexes of the rat. Chromosoma 107 540 548

30. DietrichAJ

MulderRJ

1983 A light- and electron microscopic analysis of meiotic prophase in female mice. Chromosoma 88 377 385

31. de VriesFA

de BoerE

van den BoschBM

BaarendsWM

OomsM

2005 Mouse Sycp1 functions in synaptonemal complex assembly, meiotic recombination, and XY body formation. Genes Dev 19 1376 1389

32. KeeneyS

GirouxCN

KlecknerN

1997 Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88 375 384

33. Fernandez-CapetilloO

LeeA

NussenzweigM

NussenzweigA

2004 H2AX: the histone guardian of the genome. DNA Repair 3 959 967

34. KinnerA

WuW

StaudtC

IliakisG

2008 Gamma-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids Res 36 5678 5694

35. KeeneyS

2001 Mechanism and control of meiotic recombination initiation. Curr Top Dev Biol 52 1 53

36. MahadevaiahSK

TurnerJM

BaudatF

RogakouEP

de BoerP

2001 Recombinational DNA double-strand breaks in mice precede synapsis. Nat Genet 27 271 276

37. OgawaT

YuX

ShinoharaA

EgelmanEH

1993 Similarity of the yeast RAD51 filament to the bacterial RecA filament. Science 259 1896 1899

38. TarsounasM

MoritaT

PearlmanRE

MoensPB

1999 RAD51 and DMC1 form mixed complexes associated with mouse meiotic chromosome cores and synaptonemal complexes. J Cell Biol 147 207 220

39. WuX

MondalG

WangX

WuJ

YangL

2009 Microcephalin regulates BRCA2 and Rad51-associated DNA double-strand break repair. Cancer Res 69 5531 5536

40. XuY

AshleyT

BrainerdEE

BronsonRT

MeynMS

1996 Targeted disruption of ATM leads to growth retardation, chromosomal fragmentation during meiosis, immune defects, and thymic lymphoma. Genes Dev 10 2411 2422

41. CelesteA

PetersenS

RomanienkoPJ

Fernandez-CapetilloO

ChenHT

2002 Genomic instability in mice lacking histone H2AX. Science 296 922 927

42. LouZ

Minter-DykhouseK

FrancoS

GostissaM

RiveraMA

2006 MDC1 maintains genomic stability by participating in the amplification of ATM-dependent DNA damage signals. Mol Cell 21 187 200

43. YangSZ

LinFT

LinWC

2008 MCPH1/BRIT1 cooperates with E2F1 in the activation of checkpoint, DNA repair and apoptosis. EMBO Rep 9 907 915

44. SharanSK

PyleA

CoppolaV

BabusJ

SwaminathanS

2004 BRCA2 deficiency in mice leads to meiotic impairment and infertility. Development 131 131 142

45. PittmanDL

CobbJ

SchimentiKJ

WilsonLA

CooperDM

1998 Meiotic prophase arrest with failure of chromosome synapsis in mice deficient for Dmc1, a germline-specific RecA homolog. Mol Cell 1 697 705

46. YoshidaK

KondohG

MatsudaY

HabuT

NishimuneY

1998 The mouse RecA-like gene Dmc1 is required for homologous chromosome synapsis during meiosis. Mol Cell 1 707 718

47. LiK

RamirezMA

RoseE

BeaudetAL

2002 A gene fusion method to screen for regulatory effects on gene expression: application to the LDL receptor. Hum Mol Genet 11 3257 3265

48. WardIM

MinnK

van DeursenJ

ChenJ

2003 p53 Binding protein 53BP1 is required for DNA damage responses and tumor suppression in mice. Mol Cell Biol 23 2556 2563

49. MeiJ

HuangX

ZhangP

2001 Securin is not required for cellular viability, but is required for normal growth of mouse embryonic fibroblasts. Curr Biol 11 1197 1201

50. PetersAH

PlugAW

van VugtMJ

de BoerP

1997 A drying-down technique for the spreading of mammalian meiocytes from the male and female germline. Chromosome Res 5 66 68

51. BaartEB

de RooijDG

KeeganKS

de BoerP

2000 Distribution of Atr protein in primary spermatocytes of a mouse chromosomal mutant: a comparison of preparation techniques. Chromosoma 109 139 147