The Polyproline Site in Hinge 2 Influences the Functional Capacity of Truncated Dystrophins

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Mutations in dystrophin can lead to Duchenne muscular dystrophy or the more mild form of the disease, Becker muscular dystrophy. The hinge 3 region in the rod domain of dystrophin is particularly prone to deletion mutations. In-frame deletions of hinge 3 are predicted to lead to BMD, however the severity of disease can vary considerably. Here we performed extensive structure-function analyses of truncated dystrophins with modified hinges and spectrin-like repeats in mdx mice. We found that the polyproline site in hinge 2 profoundly influences the functional capacity of a microdystrophin^{ΔR4-R23/ΔCT} with a large deletion in the hinge 3 region. Inclusion of polyproline in microdystrophin^{ΔR4-R23/ΔCT} led to small myofibers (12% smaller than wild-type), Achilles myotendinous disruption, ringed fibers, and aberrant neuromuscular junctions in the mdx gastrocnemius muscles. Replacing hinge 2 of microdystrophin^{ΔR4-R23/ΔCT} with hinge 3 significantly improved the functional capacity to prevent muscle degeneration, increase muscle fiber area, and maintain the junctions. We conclude that the rigid α-helical structure of the polyproline site significantly impairs the functional capacity of truncated dystrophins to maintain appropriate connections between the cytoskeleton and extracellular matrix.

Vyšlo v časopise: The Polyproline Site in Hinge 2 Influences the Functional Capacity of Truncated Dystrophins. PLoS Genet 6(5): e32767. doi:10.1371/journal.pgen.1000958
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1000958

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1. HoffmanEP

BrownRHJr

KunkelLM

1987 Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 51 919 928

2. KoenigM

HoffmanEP

BertelsonCJ

MonacoAP

FeenerC

1987 Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 50 509 517

3. EmeryAE

Muntoni

2003 Duchenne Muscular Dystrophy., 3rd edn. Oxford University Press, Oxford

4. BanksGB

FuhrerC

AdamsME

FroehnerSC

2003 The postsynaptic submembrane machinery at the neuromuscular junction: requirement for rapsyn and the utrophin/dystrophin-associated complex. J Neurocytol 32 709 726

5. BhasinN

LawR

LiaoG

SaferD

EllmerJ

2005 Molecular extensibility of mini-dystrophins and a dystrophin rod construct. J Mol Biol 352 795 806

6. ErvastiJM

2007 Dystrophin, its interactions with other proteins, and implications for muscular dystrophy. Biochim Biophys Acta 1772 108 117

7. ChamberlainJS

GibbsRA

RanierJE

NguyenPN

CaskeyCT

1988 Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res 16 11141 11156

8. BaumbachLL

ChamberlainJS

WardPA

FarwellNJ

CaskeyCT

1989 Molecular and clinical correlations of deletions leading to Duchenne and Becker muscular dystrophies. Neurology 39 465 474

9. GillardEF

ChamberlainJS

MurphyEG

DuffCL

SmithB

1989 Molecular and phenotypic analysis of patients with deletions within the deletion-rich region of the Duchenne muscular dystrophy (DMD) gene. Am J Hum Genet 45 507 520

10. KoenigM

BeggsAH

MoyerM

ScherpfS

HeindrichK

1989 The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet 45 498 506

11. WerneckLC

ScolaRH

MaegawaGH

WerneckMC

2001 Comparative analysis of PCR-deletion detection and immunohistochemistry in Brazilian Duchenne and Becker muscular dystrophy patients. Am J Med Genet 103 115 120

12. Den DunnenJT

GrootscholtenPM

BakkerE

BlondenLA

GinjaarHB

1989 Topography of the Duchenne muscular dystrophy (DMD) gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications. Am J Hum Genet 45 835 847

13. Coral-VazquezR

ArenasD

CisnerosB

PenalozaL

KofmanS

1993 Analysis of dystrophin gene deletions in patients from the Mexican population with Duchenne/Becker muscular dystrophy. Arch Med Res 24 1 6

14. FlaniganKM

DunnDM

von NiederhausernA

SoltanzadehP

GappmaierE

2009 Mutational spectrum of DMD mutations in dystrophinopathy patients: application of modern diagnostic techniques to a large cohort. Hum Mutat 30 1657 1666

15. AbmayrS

ChamberlainJ

2006 The structure and function of dystrophin.

WinderSJ

Molecular Mechanisms of Muscular Dystrophies Georgetown Landes Biosciences 14 34

16. BanksGB

ChamberlainJS

2008 The value of mammalian models for duchenne muscular dystrophy in developing therapeutic strategies. Curr Top Dev Biol 84 431 453

17. KoenigM

MonacoAP

KunkelLM

1988 The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell 53 219 226

18. DavisonMD

BaronMD

CritchleyDR

WoottonJC

1989 Structural analysis of homologous repeated domains in alpha-actinin and spectrin. Int J Biol Macromol 11 81 90

19. KoenigM

KunkelLM

1990 Detailed analysis of the repeat domain of dystrophin reveals four potential hinge segments that may confer flexibility. J Biol Chem 265 4560 4566

20. AmannKJ

RenleyBA

ErvastiJM

1998 A cluster of basic repeats in the dystrophin rod domain binds F-actin through an electrostatic interaction. J Biol Chem 273 28419 28423

21. CarsanaA

FrissoG

TremolaterraMR

LanzilloR

VitaleDF

2005 Analysis of dystrophin gene deletions indicates that the hinge III region of the protein correlates with disease severity. Ann Hum Genet 69 253 259

22. PhelpsSF

HauserMA

ColeNM

RafaelJA

HinkleRT

1995 Expression of full-length and truncated dystrophin mini-genes in transgenic mdx mice. Hum Mol Genet 4 1251 1258

23. RafaelJA

CoxGA

CorradoK

JungD

CampbellKP

1996 Forced expression of dystrophin deletion constructs reveals structure-function correlations. J Cell Biol 134 93 102

24. HarperSQ

HauserMA

DelloRussoC

DuanD

CrawfordRW

2002 Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy. Nat Med 8 253 261

25. CrawfordGE

FaulknerJA

CrosbieRH

CampbellKP

FroehnerSC

2000 Assembly of the dystrophin-associated protein complex does not require the dystrophin COOH-terminal domain. J Cell Biol 150 1399 1410

26. WangB

LiJ

XiaoX

2000 Adeno-associated virus vector carrying human minidystrophin genes effectively ameliorates muscular dystrophy in mdx mouse model. Proc Natl Acad Sci U S A 97 13714 13719

27. YuasaK

MiyagoeY

YamamotoK

NabeshimaY

DicksonG

1998 Effective restoration of dystrophin-associated proteins in vivo by adenovirus-mediated transfer of truncated dystrophin cDNAs. FEBS Lett 425 329 336

28. GregorevicP

BlankinshipMJ

AllenJM

CrawfordRW

MeuseL

2004 Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat Med 10 828 834

29. GregorevicP

AllenJM

MinamiE

BlankinshipMJ

HaraguchiM

2006 rAAV6-microdystrophin preserves muscle function and extends lifespan in severely dystrophic mice. Nat Med 12 787 789

30. YoshimuraM

SakamotoM

IkemotoM

MochizukiY

YuasaK

2004 AAV vector-mediated microdystrophin expression in a relatively small percentage of mdx myofibers improved the mdx phenotype. Mol Ther 10 821 828

31. YueY

LiuM

DuanD

2006 C-terminal-truncated microdystrophin recruits dystrobrevin and syntrophin to the dystrophin-associated glycoprotein complex and reduces muscular dystrophy in symptomatic utrophin/dystrophin double-knockout mice. Mol Ther 14 79 87

32. BanksGB

GregorevicP

AllenJM

FinnEE

ChamberlainJS

2007 Functional capacity of dystrophins carrying deletions in the N-terminal actin-binding domain. Hum Mol Genet 16(17) 2105 2113

33. BanksGB

CombsAC

ChamberlainJR

ChamberlainJS

2008 Molecular and cellular adaptations to chronic myotendinous strain injury in mdx mice expressing a truncated dystrophin. Hum Mol Genet (17)24 3975 3986

34. FosterH

SharpPS

AthanasopoulosT

TrolletC

GrahamIR

2008 Codon and mRNA Sequence Optimization of Microdystrophin Transgenes Improves Expression and Physiological Outcome in Dystrophic mdx Mice Following AAV2/8 Gene Transfer. Mol Ther

35. GregorevicP

BlankinshipMJ

AllenJM

ChamberlainJS

2008 Systemic microdystrophin gene delivery improves skeletal muscle structure and function in old dystrophic mdx mice. Mol Ther 16 657 664

36. BanksGB

ChamberlainJS

FroehnerSC

2009 Truncated dystrophins can influence neuromuscular synapse structure. Mol Cell Neurosci 40 433 441

37. TidballJG

1991 Force transmission across muscle cell membranes. J Biomech 24 Suppl 1 43 52

38. LyonsPR

SlaterCR

1991 Structure and function of the neuromuscular junction in young adult mdx mice. J Neurocytol 20 969 981

39. GradyRM

TengH

NicholMC

CunninghamJC

WilkinsonRS

1997 Skeletal and cardiac myopathies in mice lacking utrophin and dystrophin: a model for Duchenne muscular dystrophy. Cell 90 729 738

40. LynchGS

2004 Role of contraction-induced injury in the mechanisms of muscle damage in muscular dystrophy. Clin Exp Pharmacol Physiol 31 557 561

41. KayBK

WilliamsonMP

SudolM

2000 The importance of being proline: the interaction of proline-rich motifs in signaling proteins with their cognate domains. Faseb J 14 231 241

42. RathA

DavidsonAR

DeberCM

2005 The structure of “unstructured” regions in peptides and proteins: role of the polyproline II helix in protein folding and recognition. Biopolymers 80 179 185

43. JudgeLM

ChamberlainJS

2005 Gene therapy for Duchenne muscular dystrophy: AAV leads the way. Acta Myol 24 184 193

44. GehrigSM

KoopmanR

NaimT

TjoakarfaC

LynchGS

2009 Making Fast-Twitch Dystrophic Muscles Bigger Protects Them from Contraction Injury and Attenuates the Dystrophic Pathology. Am J Pathol

45. CrossRA

StewartM

Kendrick-JonesJ

1990 Structural predictions for the central domain of dystrophin. FEBS Lett 262 87 92

46. YanY

WinogradE

VielA

CroninT

HarrisonSC

1993 Crystal structure of the repetitive segments of spectrin. Science 262 2027 2030

47. KahanaE

MarshPJ

HenryAJ

WayM

GratzerWB

1994 conformation and phasing of dystrophin structural repeats. J Mol Biol 235 1271 1277

48. CalvertR

KahanaE

GratzerWB

1996 Stability of the dystrophin rod domain fold: evidence for nested repeating units. Biophys J 71 1605 1610

49. PascualJ

CastresanaJ

SarasteM

1997 Evolution of the spectrin repeat. Bioessays 19 811 817

50. BroderickMJ

WinderSJ

2002 Towards a complete atomic structure of spectrin family proteins. J Struct Biol 137 184 193

51. HarperSQ

CrawfordRW

DelloRussoC

ChamberlainJS

2002 Spectrin-like repeats from dystrophin and alpha-actinin-2 are not functionally interchangeable. Hum Mol Genet 11 1807 1815

52. SaadatL

PittmanL

MenhartN

2006 Structural cooperativity in spectrin type repeats motifs of dystrophin. Biochim Biophys Acta 1764 943 954

53. WinderSJ

GibsonTJ

Kendrick-JonesJ

1995 Dystrophin and utrophin: the missing links! FEBS Lett 369 27 33

54. KahanaE

GratzerWB

1995 Minimum folding unit of dystrophin rod domain. Biochemistry 34 8110 8114

55. PascualJ

PfuhlM

WaltherD

SarasteM

NilgesM

1997 Solution structure of the spectrin repeat: a left-handed antiparallel triple-helical coiled-coil. J Mol Biol 273 740 751

56. ErvastiJM

CampbellKP

1993 A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin. J Cell Biol 122 809 823

57. RybakovaIN

PatelJR

ErvastiJM

2000 The dystrophin complex forms a mechanically strong link between the sarcolemma and costameric actin. J Cell Biol 150 1209 1214

58. ErvastiJM

2003 Costameres: the Achilles' heel of Herculean muscle. J Biol Chem 278 13591 13594

59. OzawaE

2006 The functional biology of dystrophin: structural components and the pathogenesis of Duchenne muscular dystrophy.

ChamberlainJS

RandoTA

Duchenne Muscular Dystrophy: Advances in Therapeutics New York Taylor and Francis 21 54

60. MoradiM

BabinV

RolandC

DardenTA

SaguiC

2009 Conformations and free energy landscapes of polyproline peptides. Proc Natl Acad Sci U S A

61. Ishikawa-SakuraiM

YoshidaM

ImamuraM

DaviesKE

OzawaE

2004 ZZ domain is essentially required for the physiological binding of dystrophin and utrophin to beta-dystroglycan. Hum Mol Genet 13 693 702

62. LaiY

ThomasGD

YueY

YangHT

LiD

2009 Dystrophins carrying spectrin-like repeats 16 and 17 anchor nNOS to the sarcolemma and enhance exercise performance in a mouse model of muscular dystrophy. J Clin Invest 119 624 635

63. OdomGL

GregorevicP

AllenJM

FinnE

ChamberlainJS

2008 Microutrophin delivery through rAAV6 increases lifespan and improves muscle function in dystrophic dystrophin/utrophin-deficient mice. Mol Ther 16 1539 1545

64. SonnemannKJ

Heun-JohnsonH

TurnerAJ

BaltgalvisKA

LoweDA

2009 Functional substitution by TAT-utrophin in dystrophin-deficient mice. PLoS Med 6 e1000083 doi:10.1371/journal.pmed.1000083

65. ChamberlainJ

2004 PCR-mediated mutagenesis. In: Nature Encyclopedia of Life Sciences. Nature Publishing Group, London

66. PetersMF

AdamsME

FroehnerSC

1997 Differential association of syntrophin pairs with the dystrophin complex. J Cell Biol 138 81 93