Elevated Expression of the Integrin-Associated Protein PINCH Suppresses the Defects of Muscle Hypercontraction Mutants

English version České info

A variety of human diseases arise from mutations that alter muscle contraction. Evolutionary conservation allows genetic studies in Drosophila melanogaster to be used to better understand these myopathies and suggest novel therapeutic strategies. Integrin-mediated adhesion is required to support muscle structure and function, and expression of Integrin adhesive complex (IAC) proteins is modulated to adapt to varying levels of mechanical stress within muscle. Mutations in flapwing (flw), a catalytic subunit of myosin phosphatase, result in non-muscle myosin hyperphosphorylation, as well as muscle hypercontraction, defects in size, motility, muscle attachment, and subsequent larval and pupal lethality. We find that moderately elevated expression of the IAC protein PINCH significantly rescues flw phenotypes. Rescue requires PINCH be bound to its partners, Integrin-linked kinase and Ras suppressor 1. Rescue is not achieved through dephosphorylation of non-muscle myosin, suggesting a mechanism in which elevated PINCH expression strengthens integrin adhesion. In support of this, elevated expression of PINCH rescues an independent muscle hypercontraction mutant in muscle myosin heavy chain, Mhc^Samba1. By testing a panel of IAC proteins, we show specificity for PINCH expression in the rescue of hypercontraction mutants. These data are consistent with a model in which PINCH is present in limiting quantities within IACs, with increasing PINCH expression reinforcing existing adhesions or allowing for the de novo assembly of new adhesion complexes. Moreover, in myopathies that exhibit hypercontraction, strategic PINCH expression may have therapeutic potential in preserving muscle structure and function.

Vyšlo v časopise: Elevated Expression of the Integrin-Associated Protein PINCH Suppresses the Defects of Muscle Hypercontraction Mutants. PLoS Genet 9(3): e32767. doi:10.1371/journal.pgen.1003406
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003406

Souhrn

Zdroje

1. CohnRD, CampbellKP (2000) Molecular basis of muscular dystrophies. Muscle Nerve 23: 1456–1471.

2. MendellJR, BoueDR, MartinPT (2006) The congenital muscular dystrophies: recent advances and molecular insights. Pediatr Dev Pathol 9: 427–443.

3. SeidmanJG, SeidmanC (2001) The genetic basis for cardiomyopathy: from mutation identification to mechanistic paradigms. Cell 104: 557–567.

4. JacobyD, McKennaWJ (2012) Genetics of inherited cardiomyopathy. Eur Heart J 33: 296–304.

5. FerrusA, AcebesA, MarinMC, Hernandez-HernandezA (2000) A genetic approach to detect muscle protein interactions in vivo. Trends Cardiovasc Med 10: 293–298.

6. VigoreauxJO (2001) Genetics of the Drosophila flight muscle myofibril: a window into the biology of complex systems. Bioessays 23: 1047–1063.

7. BeallCJ, FyrbergE (1991) Muscle abnormalities in Drosophila melanogaster heldup mutants are caused by missing or aberrant troponin-I isoforms. J Cell Biol 114: 941–951.

8. DeakII, BellamyPR, BienzM, DubuisY, FennerE, et al. (1982) Mutations affecting the indirect flight muscles of Drosophila melanogaster. J Embryol Exp Morphol 69: 61–81.

9. MontanaES, LittletonJT (2004) Characterization of a hypercontraction-induced myopathy in Drosophila caused by mutations in Mhc. J Cell Biol 164: 1045–1054.

10. RaghavanS, WilliamsI, AslamH, ThomasD, SzoorB, et al. (2000) Protein phosphatase 1beta is required for the maintenance of muscle attachments. Curr Biol 10: 269–272.

11. VereshchaginaN, BennettD, SzoorB, KirchnerJ, GrossS, et al. (2004) The essential role of PP1beta in Drosophila is to regulate nonmuscle myosin. Mol Biol Cell 15: 4395–4405.

12. HomykTJr, EmersonCPJr (1988) Functional interactions between unlinked muscle genes within haploinsufficient regions of the Drosophila genome. Genetics 119: 105–121.

13. NongthombaU, CumminsM, ClarkS, VigoreauxJO, SparrowJC (2003) Suppression of muscle hypercontraction by mutations in the myosin heavy chain gene of Drosophila melanogaster. Genetics 164: 209–222.

14. MontanaES, LittletonJT (2006) Expression profiling of a hypercontraction-induced myopathy in Drosophila suggests a compensatory cytoskeletal remodeling response. J Biol Chem 281: 8100–8109.

15. PerkinsAD, EllisSJ, AsghariP, ShamsianA, MooreED, et al. (2010) Integrin-mediated adhesion maintains sarcomeric integrity. Dev Biol 338: 15–27.

16. Pines M, Das R, Ellis SJ, Morin A, Czerniecki S, et al.. (2012) Mechanical force regulates integrin turnover in Drosophila in vivo. Nat Cell Biol.

17. HigashibataA, SzewczykNJ, ConleyCA, Imamizo-SatoM, HigashitaniA, et al. (2006) Decreased expression of myogenic transcription factors and myosin heavy chains in Caenorhabditis elegans muscles developed during spaceflight. J Exp Biol 209: 3209–3218.

18. BloorJW, KiehartDP (2001) zipper Nonmuscle myosin-II functions downstream of PS2 integrin in Drosophila myogenesis and is necessary for myofibril formation. Dev Biol 239: 215–228.

19. KadrmasJL, SmithMA, ClarkKA, PronovostSM, MusterN, et al. (2004) The integrin effector PINCH regulates JNK activity and epithelial migration in concert with Ras suppressor 1. J Cell Biol 167: 1019–1024.

20. DoughertyGW, ChoppT, QiSM, CutlerML (2005) The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions. Exp Cell Res 306: 168–179.

21. LiF, ZhangY, WuC (1999) Integrin-linked kinase is localized to cell-matrix focal adhesions but not cell-cell adhesion sites and the focal adhesion localization of integrin-linked kinase is regulated by the PINCH-binding ANK repeats. J Cell Sci 112(Pt 24): 4589–4599.

22. FukudaT, ChenK, ShiX, WuC (2003) PINCH-1 is an obligate partner of integrin-linked kinase (ILK) functioning in cell shape modulation, motility, and survival. J Biol Chem 278: 51324–51333.

23. ClarkKA, McGrailM, BeckerleMC (2003) Analysis of PINCH function in Drosophila demonstrates its requirement in integrin-dependent cellular processes. Development 130: 2611–2621.

24. ZervasCG, GregorySL, BrownNH (2001) Drosophila integrin-linked kinase is required at sites of integrin adhesion to link the cytoskeleton to the plasma membrane. J Cell Biol 152: 1007–1018.

25. VakaloglouKM, ChountalaM, ZervasCG (2012) Functional analysis of parvin and different modes of IPP-complex assembly at integrin sites during Drosophila development. J Cell Sci

26. KogataN, TribeRM, FasslerR, WayM, AdamsRH (2009) Integrin-linked kinase controls vascular wall formation by negatively regulating Rho/ROCK-mediated vascular smooth muscle cell contraction. Genes Dev 23: 2278–2283.

27. MontanezE, WickstromSA, AltstatterJ, ChuH, FasslerR (2009) Alpha-parvin controls vascular mural cell recruitment to vessel wall by regulating RhoA/ROCK signalling. EMBO J 28: 3132–3144.

28. StanchiF, GrashoffC, Nguemeni YongaCF, GrallD, FasslerR, et al. (2009) Molecular dissection of the ILK-PINCH-parvin triad reveals a fundamental role for the ILK kinase domain in the late stages of focal-adhesion maturation. J Cell Sci 122: 1800–1811.

29. MederB, HuttnerIG, Sedaghat-HamedaniF, JustS, DahmeT, et al. (2011) PINCH proteins regulate cardiac contractility by modulating integrin-linked kinase-protein kinase B signaling. Mol Cell Biol 31: 3424–3435.

30. EliasMC, PronovostSM, CahillKJ, BeckerleMC, KadrmasJL (2012) A crucial role for Ras Suppressor-1 (RSU-1) revealed when PINCH and ILK binding is disrupted. J Cell Sci

31. EkeI, KochU, HehlgansS, SandfortV, StanchiF, et al. (2010) PINCH1 regulates Akt1 activation and enhances radioresistance by inhibiting PP1alpha. J Clin Invest 120: 2516–2527.

32. ChenK, TuY, ZhangY, BlairHC, ZhangL, et al. (2008) PINCH-1 regulates the ERK-Bim pathway and contributes to apoptosis resistance in cancer cells. J Biol Chem 283: 2508–2517.

33. XuZ, FukudaT, LiY, ZhaX, QinJ, et al. (2005) Molecular dissection of PINCH-1 reveals a mechanism of coupling and uncoupling of cell shape modulation and survival. J Biol Chem 280: 27631–27637.

34. ZhangY, ChenK, TuY, VelyvisA, YangY, et al. (2002) Assembly of the PINCH-ILK-CH-ILKBP complex precedes and is essential for localization of each component to cell-matrix adhesion sites. J Cell Sci 115: 4777–4786.

35. EliasMC, PronovostSM, CahillKJ, BeckerleMC, KadrmasJL (2012) A crucial role for Ras suppressor-1 (RSU-1) revealed when PINCH and ILK binding is disrupted. J Cell Sci 125: 3185–3194.

36. ShephardF, AdenleAA, JacobsonLA, SzewczykNJ (2011) Identification and functional clustering of genes regulating muscle protein degradation from amongst the known C. elegans muscle mutants. PLoS ONE 6: e24686 doi:10.1371/journal.pone.0024686.

37. EtheridgeT, OczypokEA, LehmannS, FieldsBD, ShephardF, et al. (2012) Calpains mediate integrin attachment complex maintenance of adult muscle in Caenorhabditis elegans. PLoS Genet 8: e1002471 doi:10.1371/journal.pgen.1002471.

38. Quinones-CoelloAT, PetrellaLN, AyersK, MelilloA, MazzalupoS, et al. (2007) Exploring strategies for protein trapping in Drosophila. Genetics 175: 1089–1104.

39. SchnorrerF, SchonbauerC, LangerCC, DietzlG, NovatchkovaM, et al. (2010) Systematic genetic analysis of muscle morphogenesis and function in Drosophila. Nature 464: 287–291.

40. LealSM, NeckameyerWS (2002) Pharmacological evidence for GABAergic regulation of specific behaviors in Drosophila melanogaster. J Neurobiol 50: 245–261.

41. GrabbeC, ZervasCG, HunterT, BrownNH, PalmerRH (2004) Focal adhesion kinase is not required for integrin function or viability in Drosophila. Development 131: 5795–5805.

42. TorglerCN, NarasimhaM, KnoxAL, ZervasCG, VernonMC, et al. (2004) Tensin stabilizes integrin adhesive contacts in Drosophila. Dev Cell 6: 357–369.

43. StanchiF, BordoyR, KudlacekO, BraunA, PfeiferA, et al. (2005) Consequences of loss of PINCH2 expression in mice. J Cell Sci 118: 5899–5910.

44. ChenH, HuangXN, YanW, ChenK, GuoL, et al. (2005) Role of the integrin-linked kinase/PINCH1/alpha-parvin complex in cardiac myocyte hypertrophy. Lab Invest 85: 1342–1356.

45. ZhangY, ChenK, GuoL, WuC (2002) Characterization of PINCH-2, a new focal adhesion protein that regulates the PINCH-1-ILK interaction, cell spreading, and migration. J Biol Chem 277: 38328–38338.

46. ZhangY, GuoL, ChenK, WuC (2002) A critical role of the PINCH-integrin-linked kinase interaction in the regulation of cell shape change and migration. J Biol Chem 277: 318–326.

47. GuoL, WuC (2002) Regulation of fibronectin matrix deposition and cell proliferation by the PINCH-ILK-CH-ILKBP complex. Faseb J 16: 1298–1300.

48. RenfranzPJ, BlankmanE, BeckerleMC (2010) The cytoskeletal regulator zyxin is required for viability in Drosophila melanogaster. Anat Rec (Hoboken) 293: 1455–1469.

49. YuanL, FairchildMJ, PerkinsAD, TanentzapfG (2010) Analysis of integrin turnover in fly myotendinous junctions. J Cell Sci 123: 939–946.

50. BecamIE, TanentzapfG, LepesantJA, BrownNH, HuynhJR (2005) Integrin-independent repression of cadherin transcription by talin during axis formation in Drosophila. Nat Cell Biol 7: 510–516.

51. KadrmasJL, SmithMA, PronovostSM, BeckerleMC (2007) Characterization of RACK1 function in Drosophila development. Dev Dyn 236: 2207–2215.

52. JamesP, HalladayJ, CraigEA (1996) Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144: 1425–1436.

53. DialynasG, FlanneryKM, ZirbelLN, NagyPL, MathewsKD, et al. (2012) LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle. Hum Mol Genet 21: 1544–1556.

54. FyrbergE, BernsteinS, VijayRaghavanS (1994) Basic Methods for Drosophila muscle biology. Methods Cell Biol 44: 237–258.