Phosphatase-Dead Myotubularin Ameliorates X-Linked Centronuclear Myopathy Phenotypes in Mice

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Myotubularin MTM1 is a phosphoinositide (PPIn) 3-phosphatase mutated in X-linked centronuclear myopathy (XLCNM; myotubular myopathy). We investigated the involvement of MTM1 enzymatic activity on XLCNM phenotypes. Exogenous expression of human MTM1 in yeast resulted in vacuolar enlargement, as a consequence of its phosphatase activity. Expression of mutants from patients with different clinical progression and determination of PtdIns3P and PtdIns5P cellular levels confirmed the link between vacuolar morphology and MTM1 phosphatase activity, and showed that some disease mutants retain phosphatase activity. Viral gene transfer of phosphatase-dead myotubularin mutants (MTM1^C375S and MTM1^S376N) significantly improved most histological signs of XLCNM displayed by a Mtm1-null mouse, at similar levels as wild-type MTM1. Moreover, the MTM1^C375S mutant improved muscle performance and restored the localization of nuclei, triad alignment, and the desmin intermediate filament network, while it did not normalize PtdIns3P levels, supporting phosphatase-independent roles of MTM1 in maintaining normal muscle performance and organelle positioning in skeletal muscle. Among the different XLCNM signs investigated, we identified only triad shape and fiber size distribution as being partially dependent on MTM1 phosphatase activity. In conclusion, this work uncovers MTM1 roles in the structural organization of muscle fibers that are independent of its enzymatic activity. This underlines that removal of enzymes should be used with care to conclude on the physiological importance of their activity.

Vyšlo v časopise: Phosphatase-Dead Myotubularin Ameliorates X-Linked Centronuclear Myopathy Phenotypes in Mice. PLoS Genet 8(10): e32767. doi:10.1371/journal.pgen.1002965
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002965

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Zdroje

1. LaporteJ, HuLJ, KretzC, MandelJL, KioschisP, et al. (1996) A gene mutated in X-linked myotubular myopathy defines a new putative tyrosine phosphatase family conserved in yeast. Nat Genet 13: 175–182.

2. JungbluthH, Wallgren-PetterssonC, LaporteJ (2008) Centronuclear (myotubular) myopathy. Orphanet J Rare Dis 3: 26.

3. BiancalanaV, CaronO, GallatiS, BaasF, KressW, et al. (2003) Characterisation of mutations in 77 patients with X-linked myotubular myopathy, including a family with a very mild phenotype. Hum Genet 112: 135–142.

4. Buj-BelloA, LaugelV, MessaddeqN, ZahreddineH, LaporteJ, et al. (2002) The lipid phosphatase myotubularin is essential for skeletal muscle maintenance but not for myogenesis in mice. Proc Natl Acad Sci U S A 99: 15060–15065.

5. LaporteJ, BiancalanaV, TannerSM, KressW, SchneiderV, et al. (2000) MTM1 mutations in X-linked myotubular myopathy. Hum Mutat 15: 393–409.

6. HermanGE, KopaczK, ZhaoW, MillsPL, MetzenbergA, et al. (2002) Characterization of mutations in fifty North American patients with X-linked myotubular myopathy. Hum Mutat 19: 114–121.

7. TsaiTC, HorinouchiH, NoguchiS, MinamiN, MurayamaK, et al. (2005) Characterization of MTM1 mutations in 31 Japanese families with myotubular myopathy, including a patient carrying 240 kb deletion in Xq28 without male hypogenitalism. Neuromuscul Disord 15: 245–252.

8. LaporteJ, KressW, MandelJL (2001) Diagnosis of X-linked myotubular myopathy by detection of myotubularin. Ann Neurol 50: 42–46.

9. ToschV, VasliN, KretzC, NicotAS, GasnierC, et al. (2010) Novel molecular diagnostic approaches for X-linked centronuclear (myotubular) myopathy reveal intronic mutations. Neuromuscul Disord 20: 375–381.

10. CaldwellKK, LipsDL, BansalVS, MajerusPW (1991) Isolation and characterization of two 3-phosphatases that hydrolyze both phosphatidylinositol 3-phosphate and inositol 1,3-bisphosphate. J Biol Chem 266: 18378–18386.

11. NandurkarHH, LaytonM, LaporteJ, SelanC, CorcoranL, et al. (2003) Identification of myotubularin as the lipid phosphatase catalytic subunit associated with the 3-phosphatase adapter protein, 3-PAP. Proc Natl Acad Sci U S A 100: 8660–8665.

12. TaylorGS, MaehamaT, DixonJE (2000) Myotubularin, a protein tyrosine phosphatase mutated in myotubular myopathy, dephosphorylates the lipid second messenger, phosphatidylinositol 3-phosphate. Proc Natl Acad Sci U S A 97: 8910–8915.

13. BlondeauF, LaporteJ, BodinS, Superti-FurgaG, PayrastreB, et al. (2000) Myotubularin, a phosphatase deficient in myotubular myopathy, acts on phosphatidylinositol 3-kinase and phosphatidylinositol 3-phosphate pathway. Hum Mol Genet 9: 2223–2229.

14. SchaletzkyJ, DoveSK, ShortB, LorenzoO, ClagueMJ, et al. (2003) Phosphatidylinositol-5-phosphate activation and conserved substrate specificity of the myotubularin phosphatidylinositol 3-phosphatases. Curr Biol 13: 504–509.

15. BackerJM (2008) The regulation and function of Class III PI3Ks: novel roles for Vps34. Biochem J 410: 1–17.

16. LecompteO, PochO, LaporteJ (2008) PtdIns5P regulation through evolution: roles in membrane trafficking? Trends Biochem Sci 30: 453–460.

17. VelichkovaM, JuanJ, KadandaleP, JeanS, RibeiroI, et al. (2010) Drosophila Mtm and class II PI3K coregulate a PI(3)P pool with cortical and endolysosomal functions. J Cell Biol 190: 407–425.

18. DoveSK, DongK, KobayashiT, WilliamsFK, MichellRH (2009) Phosphatidylinositol 3,5-bisphosphate and Fab1p/PIKfyve underPPIn endo-lysosome function. Biochem J 419: 1–13.

19. LaporteJ, BedezF, BolinoA, MandelJL (2003) Myotubularins, a large disease-associated family of cooperating catalytically active and inactive phosphoinositides phosphatases. Hum Mol Genet 12 Spec No 2: R285–292.

20. RobinsonFL, DixonJE (2006) Myotubularin phosphatases: policing 3-phosphoinositides. Trends Cell Biol 16: 403–412.

21. ParrishWR, StefanCJ, EmrSD (2004) Essential role for the myotubularin-related phosphatase Ymr1p and the synaptojanin-like phosphatases Sjl2p and Sjl3p in regulation of phosphatidylinositol 3-phosphate in yeast. Mol Biol Cell 15: 3567–3579.

22. CoxK, GattasM, HarveyP, DolphinC, FriendK, et al. (2005) X-linked myotubular myopathy: mutation R69C identified in a family with multiple neonatal deaths. Clin Genet 67: 441–442.

23. LaporteJ, LiaubetL, BlondeauF, TronchereH, MandelJL, et al. (2002) Functional redundancy in the myotubularin family. Biochem Biophys Res Commun 291: 305–312.

24. WalkerDM, UrbeS, DoveSK, TenzaD, RaposoG, et al. (2001) Characterization of MTMR3. an inositol lipid 3-phosphatase with novel substrate specificity. Curr Biol 11: 1600–1605.

25. GaryJD, WurmserAE, BonangelinoCJ, WeismanLS, EmrSD (1998) Fab1p is essential for PtdIns(3)P 5-kinase activity and the maintenance of vacuolar size and membrane homeostasis. J Cell Biol 143: 65–79.

26. DoveSK, CookeFT, DouglasMR, SayersLG, ParkerPJ, et al. (1997) Osmotic stress activates phosphatidylinositol-3,5-bisphosphate synthesis. Nature 390: 187–192.

27. BonangelinoCJ, NauJJ, DuexJE, BrinkmanM, WurmserAE, et al. (2002) Osmotic stress-induced increase of phosphatidylinositol 3,5-bisphosphate requires Vac14p, an activator of the lipid kinase Fab1p. J Cell Biol 156: 1015–1028.

28. TronchereH, LaporteJ, PendariesC, ChaussadeC, LiaubetL, et al. (2004) Production of phosphatidylinositol 5-phosphate by the phosphoinositide 3-phosphatase myotubularin in mammalian cells. J Biol Chem 279: 7304–7312.

29. MorrisJB, HinchliffeKA, CiruelaA, LetcherAJ, IrvineRF (2000) Thrombin stimulation of platelets causes an increase in phosphatidylinositol 5-phosphate revealed by mass assay. FEBS Lett 475: 57–60.

30. Al-QusairiL, WeissN, ToussaintA, BerbeyC, MessaddeqN, et al. (2009) T-tubule disorganization and defective excitation-contraction coupling in muscle fibers lacking myotubularin lipid phosphatase. Proc Natl Acad Sci U S A 106: 18763–18768.

31. Buj-BelloA, FougerousseF, SchwabY, MessaddeqN, SpehnerD, et al. (2008) AAV-mediated intramuscular delivery of myotubularin corrects the myotubular myopathy phenotype in targeted murine muscle and suggests a function in plasma membrane homeostasis. Hum Mol Genet 17: 2132–2143.

32. HniaK, TronchereH, TomczakKK, AmoasiiL, SchultzP, et al. (2011) Myotubularin controls desmin intermediate filament architecture and mitochondrial dynamics in human and mouse skeletal muscle. J Clin Invest 121: 70–85.

33. DowlingJJ, VreedeAP, LowSE, GibbsEM, KuwadaJY, et al. (2009) Loss of myotubularin function results in T-tubule disorganization in zebrafish and human myotubular myopathy. PLoS Genet 5: e1000372 doi:10.1371/journal.pgen.1000372

34. ToussaintA, CowlingBS, HniaK, MohrM, OldforsA, et al. (2011) Defects in amphiphysin 2 (BIN1) and triads in several forms of centronuclear myopathies. Acta Neuropathol 121: 253–266.

35. ChicanneG, SeverinS, BoscheronC, TerrisseAD, GratacapMP, et al. (2012) A novel mass assay to quantify the bioactive lipid PtdIns3P in various biological samples. Biochem J [Epub ahead of print]

36. LaporteJ, Guiraud-ChaumeilC, VincentMC, MandelJL, TannerSM, et al. (1997) Mutations in the MTM1 gene implicated in X-linked myotubular myopathy. ENMC International Consortium on Myotubular Myopathy. European Neuro-Muscular Center. Hum Mol Genet 6: 1505–1511.

37. BegleyMJ, TaylorGS, BrockMA, GhoshP, WoodsVL, et al. (2006) Molecular basis for substrate recognition by MTMR2, a myotubularin family phosphoinositide phosphatase. Proc Natl Acad Sci U S A 103: 927–932.

38. RibeiroI, YuanL, TanentzapfG, DowlingJJ, KigerA (2011) Phosphoinositide regulation of integrin trafficking required for muscle attachment and maintenance. PLoS Genet 7: e1001295 doi:10.1371/journal.pgen.1001295

39. PiersonCR, Dulin-SmithAN, DurbanAN, MarshallML, MarshallJT, et al. (2012) Modeling the human MTM1 p.R69C mutation in murine Mtm1 results in exon 4 skipping and a less severe myotubular myopathy phenotype. Hum Mol Genet 21: 811–825.

40. LaporteJ, BlondeauF, GansmullerA, LutzY, VoneschJL, et al. (2002) The PtdIns3P phosphatase myotubularin is a cytoplasmic protein that also localizes to Rac1-inducible plasma membrane ruffles. J Cell Sci 115: 3105–3117.

41. PatruccoE, NotteA, BarberisL, SelvetellaG, MaffeiA, et al. (2004) PI3Kgamma modulates the cardiac response to chronic pressure overload by distinct kinase-dependent and -independent effects. Cell 118: 375–387.

42. VanhaesebroeckB, RohnJL, WaterfieldMD (2004) Gene targeting: attention to detail. Cell 118: 274–276.

43. VarnaiP, BallaT (2007) Visualization and manipulation of phosphoinositide dynamics in live cells using engineered protein domains. Pflugers Arch 455: 69–82.

44. VidaTA, EmrSD (1995) A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol 128: 779–792.

45. HamaH, TakemotoJY, DeWaldDB (2000) Analysis of phosphoinositides in protein trafficking. Methods 20: 465–473.

46. PayrastreB (2004) Phosphoinositides: lipid kinases and phosphatases. Methods Mol Biol 273: 201–212.

47. BlighEG, DyerWJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917.

48. RivierF, RobertA, RoyuelaM, HugonG, Bonet-KerracheA, et al. (1999) Utrophin and dystrophin-associated glycoproteins in normal and dystrophin deficient cardiac muscle. J Muscle Res Cell Motil 20: 305–314.

49. TaylorGS, DixonJE (2001) An assay for phosphoinositide phosphatases utilizing fluorescent substrates. Anal Biochem 295: 122–126.