and but Not Interact in Genetic Models of Amyotrophic Lateral Sclerosis


Mutations in the SOD1 and TARDBP genes have been commonly identified in Amyotrophic Lateral Sclerosis (ALS). Recently, mutations in the Fused in sarcoma gene (FUS) were identified in familial (FALS) ALS cases and sporadic (SALS) patients. Similarly to TDP-43 (coded by TARDBP gene), FUS is an RNA binding protein. Using the zebrafish (Danio rerio), we examined the consequences of expressing human wild-type (WT) FUS and three ALS–related mutations, as well as their interactions with TARDBP and SOD1. Knockdown of zebrafish Fus yielded a motor phenotype that could be rescued upon co-expression of wild-type human FUS. In contrast, the two most frequent ALS–related FUS mutations, R521H and R521C, unlike S57Δ, failed to rescue the knockdown phenotype, indicating loss of function. The R521H mutation caused a toxic gain of function when expressed alone, similar to the phenotype observed upon knockdown of zebrafish Fus. This phenotype was not aggravated by co-expression of both mutant human TARDBP (G348C) and FUS (R521H) or by knockdown of both zebrafish Tardbp and Fus, consistent with a common pathogenic mechanism. We also observed that WT FUS rescued the Tardbp knockdown phenotype, but not vice versa, suggesting that TARDBP acts upstream of FUS in this pathway. In addition we observed that WT SOD1 failed to rescue the phenotype observed upon overexpression of mutant TARDBP or FUS or upon knockdown of Tardbp or Fus; similarly, WT TARDBP or FUS also failed to rescue the phenotype induced by mutant SOD1 (G93A). Finally, overexpression of mutant SOD1 exacerbated the motor phenotype caused by overexpression of mutant FUS. Together our results indicate that TARDBP and FUS act in a pathogenic pathway that is independent of SOD1.


Vyšlo v časopise: and but Not Interact in Genetic Models of Amyotrophic Lateral Sclerosis. PLoS Genet 7(8): e32767. doi:10.1371/journal.pgen.1002214
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pgen.1002214

Souhrn

Mutations in the SOD1 and TARDBP genes have been commonly identified in Amyotrophic Lateral Sclerosis (ALS). Recently, mutations in the Fused in sarcoma gene (FUS) were identified in familial (FALS) ALS cases and sporadic (SALS) patients. Similarly to TDP-43 (coded by TARDBP gene), FUS is an RNA binding protein. Using the zebrafish (Danio rerio), we examined the consequences of expressing human wild-type (WT) FUS and three ALS–related mutations, as well as their interactions with TARDBP and SOD1. Knockdown of zebrafish Fus yielded a motor phenotype that could be rescued upon co-expression of wild-type human FUS. In contrast, the two most frequent ALS–related FUS mutations, R521H and R521C, unlike S57Δ, failed to rescue the knockdown phenotype, indicating loss of function. The R521H mutation caused a toxic gain of function when expressed alone, similar to the phenotype observed upon knockdown of zebrafish Fus. This phenotype was not aggravated by co-expression of both mutant human TARDBP (G348C) and FUS (R521H) or by knockdown of both zebrafish Tardbp and Fus, consistent with a common pathogenic mechanism. We also observed that WT FUS rescued the Tardbp knockdown phenotype, but not vice versa, suggesting that TARDBP acts upstream of FUS in this pathway. In addition we observed that WT SOD1 failed to rescue the phenotype observed upon overexpression of mutant TARDBP or FUS or upon knockdown of Tardbp or Fus; similarly, WT TARDBP or FUS also failed to rescue the phenotype induced by mutant SOD1 (G93A). Finally, overexpression of mutant SOD1 exacerbated the motor phenotype caused by overexpression of mutant FUS. Together our results indicate that TARDBP and FUS act in a pathogenic pathway that is independent of SOD1.


Zdroje

1. NelsonLM 1995 Epidemiology of ALS. Clin Neurosci 3 327 331

2. BoilleeSVande VeldeCClevelandDW 2006 ALS: a disease of motor neurons and their nonneuronal neighbors. Neuron 52 39 59

3. Gros-LouisFGasparCRouleauGA 2006 Genetics of familial and sporadic amyotrophic lateral sclerosis. Biochim Biophys Acta

4. PasinelliPBrownRH 2006 Molecular biology of amyotrophic lateral sclerosis: insights from genetics. Nat Rev Neurosci 7 710 723

5. RosenDR 1993 Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 364 362

6. KabashiEValdmanisPNDionPRouleauGA 2007 Oxidized/misfolded superoxide dismutase-1: the cause of all amyotrophic lateral sclerosis? Ann Neurol 62 553 559

7. BendottiCCarriMT 2004 Lessons from models of SOD1-linked familial ALS. Trends Mol Med 10 393 400

8. LemmensRVan HoeckeAHersmusNGeelenVD'HollanderI 2007 Overexpression of mutant superoxide dismutase 1 causes a motor axonopathy in the zebrafish. Hum Mol Genet 16 2359 2365

9. KabashiEValdmanisPNDionPSpiegelmanDMcConkeyBJ 2008 TARDBP mutations in individuals with sporadic and familial amyotrophic lateral sclerosis. Nat Genet 40 572 574

10. SreedharanJBlairIPTripathiVBHuXVanceC 2008 TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis. Science 319 1668 1672

11. Lagier-TourenneCClevelandDW 2009 Rethinking ALS: the FUS about TDP-43. Cell 136 1001 1004

12. GitchoMABalohRHChakravertySMayoKNortonJB 2008 TDP-43 A315T mutation in familial motor neuron disease. Ann Neurol 63 535 538

13. VanceCRogeljBHortobagyiTDe VosKJNishimuraAL 2009 Mutations in FUS, an RNA processing protein, cause familial amyotrophic lateral sclerosis type 6. Science 323 1208 1211

14. KwiatkowskiTJJrBoscoDALeclercALTamrazianEVanderburgCR 2009 Mutations in the FUS/TLS gene on chromosome 16 cause familial amyotrophic lateral sclerosis. Science 323 1205 1208

15. Lagier-TourenneCPolymenidouMClevelandDW TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration. Hum Mol Genet 19 R46 64

16. MackenzieIRRademakersRNeumannM TDP-43 and FUS in amyotrophic lateral sclerosis and frontotemporal dementia. Lancet Neurol 9 995 1007

17. BlairIPWilliamsKLWarraichSTDurnallJCThoengAD 2009 FUS mutations in amyotrophic lateral sclerosis: clinical, pathological, neurophysiological and genetic analysis. J Neurol Neurosurg Psychiatry

18. CorradoLDel BoRCastellottiBRattiACeredaC 2009 Mutations of FUS Gene in Sporadic Amyotrophic Lateral Sclerosis. J Med Genet

19. BelzilVVValdmanisPNDionPADaoudHKabashiE 2009 Mutations in FUS cause FALS and SALS in French and French Canadian populations. Neurology 73 1176 1179

20. ChioARestagnoGBrunettiMOssolaICalvoA 2009 Two Italian kindreds with familial amyotrophic lateral sclerosis due to FUS mutation. Neurobiol Aging 30 1272 1275

21. TicozziNSilaniVLeClercALKeaglePGelleraC 2009 Analysis of FUS gene mutation in familial amyotrophic lateral sclerosis within an Italian cohort. Neurology 73 1180 1185

22. ChioACalvoAMogliaCOssolaIBrunettiM 2010 A de novo missense mutation of the FUS gene in a “true” sporadic ALS case. Neurobiol Aging

23. YanJDengHXSiddiqueNFectoFChenW 2010 Frameshift and novel mutations in FUS in familial amyotrophic lateral sclerosis and ALS/dementia. Neurology 75 807 814

24. MillecampsSSalachasFCazeneuveCGordonPBrickaB 2010 SOD1, ANG, VAPB, TARDBP, and FUS mutations in familial amyotrophic lateral sclerosis: genotype-phenotype correlations. J Med Genet 47 554 560

25. RademakersRStewartHDejesus-HernandezMKriegerCGraff-RadfordN 2010 Fus gene mutations in familial and sporadic amyotrophic lateral sclerosis. Muscle Nerve 42 170 176

26. TateishiTHokonoharaTYamasakiRMiuraSKikuchiH 2010 Multiple system degeneration with basophilic inclusions in Japanese ALS patients with FUS mutation. Acta Neuropathol 119 355 364

27. SuzukiNAokiMWaritaHKatoMMizunoH 2010 FALS with FUS mutation in Japan, with early onset, rapid progress and basophilic inclusion. J Hum Genet 55 252 254

28. MackenzieIRRademakersRNeumannM 2010 TDP-43 and FUS in amyotrophic lateral sclerosis and frontotemporal dementia. Lancet Neurol 9 995 1007

29. BenajibaLLe BerICamuzatALacosteMThomas-AnterionC 2009 TARDBP mutations in motoneuron disease with frontotemporal lobar degeneration. Ann Neurol 65 470 473

30. BorroniBBonviciniCAlbericiABurattiEAgostiC 2009 Mutation within TARDBP leads to frontotemporal dementia without motor neuron disease. Hum Mutat 30 E974 983

31. SleegersKCrutsMVan BroeckhovenC 2010 Molecular pathways of frontotemporal lobar degeneration. Annu Rev Neurosci 33 71 88

32. Van LangenhoveTvan der ZeeJSleegersKEngelborghsSVandenbergheR 2010 Genetic contribution of FUS to frontotemporal lobar degeneration. Neurology 74 366 371

33. DengHXZhaiHBigioEHYanJFectoF FUS-immunoreactive inclusions are a common feature in sporadic and non-SOD1 familial amyotrophic lateral sclerosis. Ann Neurol 67 739 748

34. MackenzieIRBigioEHIncePGGeserFNeumannM 2007 Pathological TDP-43 distinguishes sporadic amyotrophic lateral sclerosis from amyotrophic lateral sclerosis with SOD1 mutations. Ann Neurol 61 427 434

35. LingSCAlbuquerqueCPHanJSLagier-TourenneCTokunagaS ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS. Proc Natl Acad Sci U S A

36. KimSHShanwareNPBowlerMJTibbettsRS 2010 Amyotrophic lateral sclerosis-associated proteins TDP-43 and FUS/TLS function in a common biochemical complex to co-regulate HDAC6 mRNA. J Biol Chem 285 34097 34105

37. LansonNAJrMaltareAKingHSmithRKimJH 2011 A Drosophila model of FUS-related neurodegeneration reveals genetic interaction between FUS and TDP-43. Hum Mol Genet

38. FieselFCVoigtAWeberSSVan den HauteCWaldenmaierA 2010 Knockdown of transactive response DNA-binding protein (TDP-43) downregulates histone deacetylase 6. EMBO J 29 209 221

39. PolymenidouMLagier-TourenneCHuttKRHuelgaSCMoranJ 2011 Long pre-mRNA depletion and RNA missplicing contribute to neuronal vulnerability from loss of TDP-43. Nat Neurosci 14 459 468

40. AyalaYMZagoPD'AmbrogioAXuYFPetrucelliL 2008 Structural determinants of the cellular localization and shuttling of TDP-43. J Cell Sci 121 3778 3785

41. BurattiEBaralleFE 2010 The multiple roles of TDP-43 in pre-mRNA processing and gene expression regulation. RNA Biol 7 420 429

42. ParkJLeeSBLeeSKimYSongS 2006 Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin. Nature 441 1157 1161

43. PallanckLGreenamyreJT 2006 Neurodegenerative disease: pink, parkin and the brain. Nature 441 1058

44. ValenteEMAbou-SleimanPMCaputoVMuqitMMHarveyK 2004 Hereditary early-onset Parkinson's disease caused by mutations in PINK1. Science 304 1158 1160

45. KitadaTAsakawaSHattoriNMatsumineHYamamuraY 1998 Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature 392 605 608

46. EldenACKimHJHartMPChen-PlotkinASJohnsonBS Ataxin-2 intermediate-length polyglutamine expansions are associated with increased risk for ALS. Nature 466 1069 1075

47. ValdmanisPNMeijerIAReynoldsALeiAMacLeodP 2007 Mutations in the KIAA0196 gene at the SPG8 locus cause hereditary spastic paraplegia. Am J Hum Genet 80 152 161

48. LairdASVan HoeckeADe MuynckLTimmersMVan den BoschL Progranulin is neurotrophic in vivo and protects against a mutant TDP-43 induced axonopathy. PLoS ONE 5 e13368 doi:10.1371/journal.pone.0013368

49. Gros-LouisFKrizJKabashiEMcDearmidJMillecampsS 2008 Als2 mRNA splicing variants detected in KO mice rescue severe motor dysfunction phenotype in Als2 knock-down zebrafish. Hum Mol Genet 17 2691 2702

50. KabashiELinLTradewellMLDionPABercierV 2010 Gain and loss of function of ALS-related mutations of TARDBP (TDP-43) cause motor deficits in vivo. Hum Mol Genet 19 671 683

51. SimpsonCLLemmensRMiskiewiczKBroomWJHansenVK 2009 Variants of the elongator protein 3 (ELP3) gene are associated with motor neuron degeneration. Hum Mol Genet 18 472 481

52. RameshTLyonANPinedaRHWangCJanssenPM A genetic model of amyotrophic lateral sclerosis in zebrafish displays phenotypic hallmarks of motoneuron disease. Dis Model Mech 3 652 662

53. KabashiEChampagneNBrusteinEDrapeauP In the swim of things: recent insights to neurogenetic disorders from zebrafish. Trends Genet

54. ItoDSekiMTsunodaYUchiyamaHSuzukiN 2011 Nuclear transport impairment of amyotrophic lateral sclerosis-linked mutations in FUS/TLS. Ann Neurol 69 152 162

55. GalJZhangJKwinterDMZhaiJJiaH 2010 Nuclear localization sequence of FUS and induction of stress granules by ALS mutants. Neurobiol Aging

56. TanAYManleyJL 2009 The TET family of proteins: functions and roles in disease. J Mol Cell Biol 1 82 92

57. TicozziNVanceCLeclercALKeaglePGlassJD 2011 Mutational analysis reveals the FUS homolog TAF15 as a candidate gene for familial amyotrophic lateral sclerosis. Am J Med Genet B Neuropsychiatr Genet

58. ReaumeAGElliottJLHoffmanEKKowallNWFerranteRJ 1996 Motor neurons in Cu/Zn superoxide dismutase-deficient mice develop normally but exhibit enhanced cell death after axonal injury. Nat Genet 13 43 47

59. BoscoDALemayNKoHKZhouHBurkeC Mutant FUS proteins that cause amyotrophic lateral sclerosis incorporate into stress granules. Hum Mol Genet 19 4160 4175

60. BarmadaSJSkibinskiGKorbERaoEJWuJY Cytoplasmic mislocalization of TDP-43 is toxic to neurons and enhanced by a mutation associated with familial amyotrophic lateral sclerosis. J Neurosci 30 639 649

61. DormannDRoddeREdbauerDBentmannEFischerI ALS-associated fused in sarcoma (FUS) mutations disrupt Transportin-mediated nuclear import. EMBO J 29 2841 2857

62. Liu-YesucevitzLBilgutayAZhangYJVanderwydeTCitroA Tar DNA binding protein-43 (TDP-43) associates with stress granules: analysis of cultured cells and pathological brain tissue. PLoS ONE 5 e3250 doi:10.1371/journal.pone.0013250

63. ColombritaCZennaroEFalliniCWeberMSommacalA 2009 TDP-43 is recruited to stress granules in conditions of oxidative insult. J Neurochem 111 1051 1061

64. KryndushkinDWicknerRBShewmakerF FUS/TLS forms cytoplasmic aggregates, inhibits cell growth and interacts with TDP-43 in a yeast model of amyotrophic lateral sclerosis. Protein Cell 2 223 236

65. FushimiKLongCJayaramNChenXLiL Expression of human FUS/TLS in yeast leads to protein aggregation and cytotoxicity, recapitulating key features of FUS proteinopathy. Protein Cell 2 141 149

66. Lagier-TourenneCPolymenidouMClevelandDW 2010 TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration. Hum Mol Genet 19 R46 64

67. ZhangYJXuYFDickeyCABurattiEBaralleF 2007 Progranulin mediates caspase-dependent cleavage of TAR DNA binding protein-43. J Neurosci 27 10530 10534

68. RademakersREriksenJLBakerMRobinsonTAhmedZ 2008 Common variation in the miR-659 binding-site of GRN is a major risk factor for TDP43-positive frontotemporal dementia. Hum Mol Genet 17 3631 3642

69. KleinbergerGWilsHPonsaertsPJorisGTimmermansJP 2010 Increased caspase activation and decreased TDP-43 solubility in progranulin knockout cortical cultures. J Neurochem 115 735 747

70. RitsonGPCusterSKFreibaumBDGuintoJBGeffelD TDP-43 mediates degeneration in a novel Drosophila model of disease caused by mutations in VCP/p97. J Neurosci 30 7729 7739

71. GitchoMAStriderJCarterDTaylor-ReinwaldLFormanMS 2009 VCP mutations causing frontotemporal lobar degeneration disrupt localization of TDP-43 and induce cell death. J Biol Chem 284 12384 12398

72. JohnsonJOMandrioliJBenatarMAbramzonYVan DeerlinVM 2010 Exome Sequencing Reveals VCP Mutations as a Cause of Familial ALS. Neuron 68 857 864

73. JowettT 2001 Double in situ hybridization techniques in zebrafish. Methods 23 345 358

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