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

被引:67
作者
Banks, Glen B. [1 ]
Judge, Luke M. [1 ]
Allen, James M. [1 ,2 ]
Chamberlain, Jeffrey S. [1 ,2 ,3 ]
机构
[1] Univ Washington, Senator Paul Wellstone Muscular Dystrophy Coopera, Dept Neurol, Seattle, WA 98195 USA
[2] Univ Washington, Dept Med, Seattle, WA 98195 USA
[3] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
基金
美国国家卫生研究院; 英国医学研究理事会;
关键词
DUCHENNE MUSCULAR-DYSTROPHY; SPECTRIN-LIKE REPEATS; MDX MICE; ROD DOMAIN; DMD GENE; NEUROMUSCULAR-JUNCTION; GLYCOPROTEIN COMPLEX; MUSCLE FUNCTION; MOUSE MODEL; LIFE-SPAN;
D O I
10.1371/journal.pgen.1000958
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
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(Delta R4-R23/Delta CT) with a large deletion in the hinge 3 region. Inclusion of polyproline in microdystrophin(Delta R4-R23/Delta CT) 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(Delta R4-R23/Delta CT) 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 a-helical structure of the polyproline site significantly impairs the functional capacity of truncated dystrophins to maintain appropriate connections between the cytoskeleton and extracellular matrix.
引用
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页数:10
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