Mislocalization of XPF-ERCC1 Nuclease Contributes to Reduced DNA Repair in XP-F Patients

English version České info

Xeroderma pigmentosum (XP) is caused by defects in the nucleotide excision repair (NER) pathway. NER removes helix-distorting DNA lesions, such as UV–induced photodimers, from the genome. Patients suffering from XP exhibit exquisite sun sensitivity, high incidence of skin cancer, and in some cases neurodegeneration. The severity of XP varies tremendously depending upon which NER gene is mutated and how severely the mutation affects DNA repair capacity. XPF-ERCC1 is a structure-specific endonuclease essential for incising the damaged strand of DNA in NER. Missense mutations in XPF can result not only in XP, but also XPF-ERCC1 (XFE) progeroid syndrome, a disease of accelerated aging. In an attempt to determine how mutations in XPF can lead to such diverse symptoms, the effects of a progeria-causing mutation (XPF^R153P) were compared to an XP–causing mutation (XPF^R799W) in vitro and in vivo. Recombinant XPF harboring either mutation was purified in a complex with ERCC1 and tested for its ability to incise a stem-loop structure in vitro. Both mutant complexes nicked the substrate indicating that neither mutation obviates catalytic activity of the nuclease. Surprisingly, differential immunostaining and fractionation of cells from an XFE progeroid patient revealed that XPF-ERCC1 is abundant in the cytoplasm. This was confirmed by fluorescent detection of XPF^R153P-YFP expressed in Xpf mutant cells. In addition, microinjection of XPF^R153P-ERCC1 into the nucleus of XPF–deficient human cells restored nucleotide excision repair of UV–induced DNA damage. Intriguingly, in all XPF mutant cell lines examined, XPF-ERCC1 was detected in the cytoplasm of a fraction of cells. This demonstrates that at least part of the DNA repair defect and symptoms associated with mutations in XPF are due to mislocalization of XPF-ERCC1 into the cytoplasm of cells, likely due to protein misfolding. Analysis of these patient cells therefore reveals a novel mechanism to potentially regulate a cell's capacity for DNA repair: by manipulating nuclear localization of XPF-ERCC1.

Vyšlo v časopise: Mislocalization of XPF-ERCC1 Nuclease Contributes to Reduced DNA Repair in XP-F Patients. PLoS Genet 6(3): e32767. doi:10.1371/journal.pgen.1000871
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1000871

Souhrn

Zdroje

1. KraemerKH

LeeMM

ScottoJ

1987 Xeroderma pigmentosum. Cutaneous, ocular, and neurologic abnormalities in 830 published cases. Arch Dermatol 123 241 250

2. NouspikelT

2008 Nucleotide excision repair and neurological diseases. DNA Repair (Amst) 7 1155 1167

3. RobbinsJH

BrumbackRA

MendionesM

BarrettSF

CarlJR

1991 Neurological disease in xeroderma pigmentosum. Documentation of a late onset type of the juvenile onset form. Brain 114 ( Pt 3) 1335 1361

4. GilletLC

ScharerOD

2006 Molecular mechanisms of mammalian global genome nucleotide excision repair. Chem Rev 106 253 276

5. SugasawaK

NgJM

MasutaniC

IwaiS

van der SpekPJ

1998 Xeroderma pigmentosum group C protein complex is the initiator of global genome nucleotide excision repair. Mol Cell 2 223 232

6. VolkerM

MoneMJ

KarmakarP

van HoffenA

SchulW

2001 Sequential assembly of the nucleotide excision repair factors in vivo. Mol Cell 8 213 224

7. SugasawaK

2006 UV-induced ubiquitylation of XPC complex, the UV-DDB-ubiquitin ligase complex, and DNA repair. J Mol Histol 37 189 202

8. LaineJP

EglyJM

2006 When transcription and repair meet: a complex system. Trends Genet 22 430 436

9. HanawaltPC

SpivakG

2008 Transcription-coupled DNA repair: two decades of progress and surprises. Nat Rev Mol Cell Biol 9 958 970

10. TroelstraC

van GoolA

de WitJ

VermeulenW

BootsmaD

1992 ERCC6, a member of a subfamily of putative helicases, is involved in Cockayne's syndrome and preferential repair of active genes. Cell 71 939 953

11. HenningKA

LiL

IyerN

McDanielLD

ReaganMS

1995 The Cockayne syndrome group A gene encodes a WD repeat protein that interacts with CSB protein and a subunit of RNA polymerase II TFIIH. Cell 82 555 564

12. NakatsuY

AsahinaH

CitterioE

RademakersS

VermeulenW

2000 XAB2, a novel tetratricopeptide repeat protein involved in transcription-coupled DNA repair and transcription. J Biol Chem 275 34931 34937

13. EvansE

MoggsJG

HwangJR

EglyJM

WoodRD

1997 Mechanism of open complex and dual incision formation by human nucleotide excision repair factors. Embo J 16 6559 6573

14. O'DonovanA

DaviesAA

MoggsJG

WestSC

WoodRD

1994 XPG endonuclease makes the 3′ incision in human DNA nucleotide excision repair. Nature 371 432 435

15. BardwellAJ

BardwellL

TomkinsonAE

FriedbergEC

1994 Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease. Science 265 2082 2085

16. SijbersAM

de LaatWL

ArizaRR

BiggerstaffM

WeiYF

1996 Xeroderma pigmentosum group F caused by a defect in a structure-specific DNA repair endonuclease. Cell 86 811 822

17. StaresincicL

FagbemiAF

EnzlinJH

GourdinAM

WijgersN

2009 Coordination of dual incision and repair synthesis in human nucleotide excision repair. EMBO J 28 1111 1120

18. ShivjiMK

PodustVN

HubscherU

WoodRD

1995 Nucleotide excision repair DNA synthesis by DNA polymerase epsilon in the presence of PCNA, RFC, and RPA. Biochemistry 34 5011 5017

19. MoserJ

KoolH

GiakzidisI

CaldecottK

MullendersLH

2007 Sealing of chromosomal DNA nicks during nucleotide excision repair requires XRCC1 and DNA ligase III alpha in a cell-cycle-specific manner. Mol Cell 27 311 323

20. OgiT

LehmannAR

2006 The Y-family DNA polymerase kappa (pol kappa) functions in mammalian nucleotide-excision repair. Nat Cell Biol 8 640 642

21. de LaatWL

AppeldoornE

JaspersNG

HoeijmakersJH

1998 DNA structural elements required for ERCC1-XPF endonuclease activity. J Biol Chem 273 7835 7842

22. NiedernhoferLJ

OdijkH

BudzowskaM

van DrunenE

MaasA

2004 The structure-specific endonuclease Ercc1-Xpf is required to resolve DNA interstrand cross-link-induced double-strand breaks. Mol Cell Biol 24 5776 5787

23. AhmadA

RobinsonAR

DuensingA

van DrunenE

BeverlooHB

2008 ERCC1-XPF endonuclease facilitates DNA double-strand break repair. Mol Cell Biol 28 5082 5092

24. GaillardPH

WoodRD

2001 Activity of individual ERCC1 and XPF subunits in DNA nucleotide excision repair. Nucleic Acids Res 29 872 879

25. de LaatWL

SijbersAM

OdijkH

JaspersNG

HoeijmakersJH

1998 Mapping of interaction domains between human repair proteins ERCC1 and XPF. Nucleic Acids Res 26 4146 4152

26. BiggerstaffM

SzymkowskiDE

WoodRD

1993 Co-correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro. EMBO J 12 3685 3692

27. EnzlinJH

ScharerOD

2002 The active site of the DNA repair endonuclease XPF-ERCC1 forms a highly conserved nuclease motif. Embo J 21 2045 2053

28. TsodikovOV

IvanovD

OrelliB

StaresincicL

ShoshaniI

2007 Structural basis for the recruitment of ERCC1-XPF to nucleotide excision repair complexes by XPA. EMBO J 26 4768 4776

29. LiL

ElledgeSJ

PetersonCA

BalesES

LegerskiRJ

1994 Specific association between the human DNA repair proteins XPA and ERCC1. Proc Natl Acad Sci U S A 91 5012 5016

30. ZengL

QuillietX

Chevallier-LagenteO

EvenoE

SarasinA

1997 Retrovirus-mediated gene transfer corrects DNA repair defect of xeroderma pigmentosum cells of complementation groups A, B and C. 4 Gene Ther 1077 1084

31. MoriwakiS

KraemerKH

2001 Xeroderma pigmentosum–bridging a gap between clinic and laboratory. Photodermatol Photoimmunol Photomed 17 47 54

32. NiedernhoferLJ

GarinisGA

RaamsA

LalaiAS

RobinsonAR

2006 A new progeroid syndrome reveals that genotoxic stress suppresses the somatotroph axis. Nature 444 1038 1043

33. SijbersAM

van Voorst VaderPC

SnoekJW

RaamsA

JaspersNG

1998 Homozygous R788W point mutation in the XPF gene of a patient with xeroderma pigmentosum and late-onset neurologic disease. J Invest Dermatol 110 832 836

34. VermeulenW

BergmannE

AuriolJ

RademakersS

FritP

2000 Sublimiting concentration of TFIIH transcription/DNA repair factor causes TTD-A trichothiodystrophy disorder. Nat Genet 26 307 313

35. HoeijmakersJH

EkerAP

WoodRD

RobinsP

1990 Use of in vivo and in vitro assays for the characterization of mammalian excision repair and isolation of repair proteins. Mutat Res 236 223 238

36. TaylorEM

BroughtonBC

BottaE

StefaniniM

SarasinA

1997 Xeroderma pigmentosum and trichothiodystrophy are associated with different mutations in the XPD (ERCC2) repair/transcription gene. Proc Natl Acad Sci U S A 94 8658 8663

37. VermeulenW

ScottRJ

RodgersS

MullerHJ

ColeJ

1994 Clinical heterogeneity within xeroderma pigmentosum associated with mutations in the DNA repair and transcription gene ERCC3. Am J Hum Genet 54 191 200

38. NouspikelT

LalleP

LeadonSA

CooperPK

ClarksonSG

1997 A common mutational pattern in Cockayne syndrome patients from xeroderma pigmentosum group G: implications for a second XPG function. Proc Natl Acad Sci U S A 94 3116 3121

39. ItoS

KuraokaI

ChymkowitchP

CompeE

TakedachiA

2007 XPG stabilizes TFIIH, allowing transactivation of nuclear receptors: implications for Cockayne syndrome in XP-G/CS patients. Mol Cell 26 231 243

40. KraemerKH

PatronasNJ

SchiffmannR

BrooksBP

TamuraD

2007 Xeroderma pigmentosum, trichothiodystrophy and Cockayne syndrome: a complex genotype-phenotype relationship. Neuroscience 145 1388 1396

41. de BoerJ

AndressooJO

de WitJ

HuijmansJ

BeemsRB

2002 Premature aging in mice deficient in DNA repair and transcription. Science 296 1276 1279

42. TakayamaK

SalazarEP

BroughtonBC

LehmannAR

SarasinA

1996 Defects in the DNA repair and transcription gene ERCC2(XPD) in trichothiodystrophy. Am J Hum Genet 58 263 270

43. van GoolAJ

CitterioE

RademakersS

van OsR

VermeulenW

1997 The Cockayne syndrome B protein, involved in transcription-coupled DNA repair, resides in an RNA polymerase II-containing complex. Embo J 16 5955 5965

44. ClarksonSG

2003 The XPG story. Biochimie 85 1113 1121

45. DubaeleS

Proietti De SantisL

BienstockRJ

KerielA

StefaniniM

2003 Basal transcription defect discriminates between xeroderma pigmentosum and trichothiodystrophy in XPD patients. Mol Cell 11 1635 1646

46. MatsumuraY

NishigoriC

YagiT

ImamuraS

TakebeH

1998 Characterization of molecular defects in xeroderma pigmentosum group F in relation to its clinically mild symptoms. Hum Mol Genet 7 969 974

47. ZelleB

BerendsF

LohmanPH

1980 Repair of damage by ultraviolet radiation in xeroderma pigmentosum cell strains of complementation groups E and F. Mutat Res 73 157 169

48. JaspersNG

RaamsA

SilengoMC

WijgersN

NiedernhoferLJ

2007 First reported patient with human ERCC1 deficiency has cerebro-oculo-facio-skeletal syndrome with a mild defect in nucleotide excision repair and severe developmental failure. Am J Hum Genet 80 457 466

49. MollUM

RiouG

LevineAJ

1992 Two distinct mechanisms alter p53 in breast cancer: mutation and nuclear exclusion. Proc Natl Acad Sci U S A 89 7262 7266

50. KauTR

WayJC

SilverPA

2004 Nuclear transport and cancer: from mechanism to intervention. Nat Rev Cancer 4 106 117

51. MinYH

CheongJW

KimJY

EomJI

LeeST

2004 Cytoplasmic mislocalization of p27Kip1 protein is associated with constitutive phosphorylation of Akt or protein kinase B and poor prognosis in acute myelogenous leukemia. Cancer Res 64 5225 5231

52. Rosin-ArbesfeldR

CliffeA

BrabletzT

BienzM

2003 Nuclear export of the APC tumour suppressor controls beta-catenin function in transcription. Embo J 22 1101 1113

53. KarinM

CaoY

GretenFR

LiZW

2002 NF-kappaB in cancer: from innocent bystander to major culprit. Nat Rev Cancer 2 301 310

54. FabbroM

RodriguezJA

BaerR

HendersonBR

2002 BARD1 induces BRCA1 intranuclear foci formation by increasing RING-dependent BRCA1 nuclear import and inhibiting BRCA1 nuclear export. J Biol Chem 277 21315 21324

55. EdwardsSW

TanCM

LimbirdLE

2000 Localization of G-protein-coupled receptors in health and disease. Trends Pharmacol Sci 21 304 308

56. WelshMJ

SmithAE

1993 Molecular mechanisms of CFTR chloride channel dysfunction in cystic fibrosis. Cell 73 1251 1254

57. WeedaG

DonkerI

de WitJ

MorreauH

JanssensR

1997 Disruption of mouse ERCC1 results in a novel repair syndrome with growth failure, nuclear abnormalities and senescence. Curr Biol 7 427 439

58. DolleME

BusuttilRA

GarciaAM

WijnhovenS

van DrunenE

2006 Increased genomic instability is not a prerequisite for shortened lifespan in DNA repair deficient mice. Mutat Res 596 22 35

59. PootM

VerkerkA

KosterJF

EsterbauerH

JongkindJF

1987 Influence of cumene hydroperoxide and 4-hydroxynonenal on the glutathione metabolism during in vitro ageing of human skin fibroblasts. Eur J Biochem 162 287 291

60. AraseS

KozukaT

TanakaK

IkenagaM

TakebeH

1979 A sixth complementation group in xeroderma pigmentosum. Mutat Res 59 143 146

61. BerneburgM

ClingenPH

HarcourtSA

LoweJE

TaylorEM

2000 The cancer-free phenotype in trichothiodystrophy is unrelated to its repair defect. Cancer Res 60 431 438

62. LimsirichaikulS

NiimiA

FawcettH

LehmannA

YamashitaS

2009 A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU). Nucleic Acids Res 37 e31

63. van VuurenAJ

AppeldoornE

OdijkH

HumbertS

MoncollinV

1995 Partial characterization of the DNA repair protein complex, containing the ERCC1, ERCC4, ERCC11 and XPF correcting activities. Mutat Res 337 25 39

64. KapetanakiMG

Guerrero-SantoroJ

BisiDC

HsiehCL

Rapic-OtrinV

2006 The DDB1-CUL4ADDB2 ubiquitin ligase is deficient in xeroderma pigmentosum group E and targets histone H2A at UV-damaged DNA sites. Proc Natl Acad Sci U S A 103 2588 2593

65. de JongeAJ

VermeulenW

KleinB

HoeijmakersJH

1983 Microinjection of human cell extracts corrects xeroderma pigmentosum defect. Embo J 2 637 641

66. WeedaG

van HamRC

VermeulenW

BootsmaD

van der EbAJ

1990 A presumed DNA helicase encoded by ERCC-3 is involved in the human repair disorders xeroderma pigmentosum and Cockayne's syndrome. Cell 62 777 791