title: Calpain-mediated proteolysis of tropomodulin isoforms leads to thin filament elongation in dystrophic skeletal muscle.
abstract: Duchenne muscular dystrophy (DMD) induces sarcolemmal mechanical instability and rupture, hyperactivity of intracellular calpains, and proteolytic breakdown of muscle structural proteins. Here we identify the two sarcomeric tropomodulin (Tmod) isoforms, Tmod1 and Tmod4, as novel proteolytic targets of m-calpain, with Tmod1 exhibiting ∼10-fold greater sensitivity to calpain-mediated cleavage than Tmod4 in situ. In mdx mice, increased m-calpain levels in dystrophic soleus muscle are associated with loss of Tmod1 from the thin filament pointed ends, resulting in ∼11% increase in thin filament lengths. In mdx/mTR mice, a more severe model of DMD, Tmod1 disappears from the thin filament pointed ends in both tibialis anterior (TA) and soleus muscles, whereas Tmod4 additionally disappears from soleus muscle, resulting in thin filament length increases of ∼10 and ∼12% in TA and soleus muscles, respectively. In both mdx and mdx/mTR mice, both TA and soleus muscles exhibit normal localization of α-actinin, the nebulin M1M2M3 domain, Tmod3, and cytoplasmic γ-actin, indicating that m-calpain does not cause wholesale proteolysis of other sarcomeric and actin cytoskeletal proteins in dystrophic skeletal muscle. These results implicate Tmod proteolysis and resultant thin filament length misspecification as novel mechanisms that may contribute to DMD pathology, affecting muscles in a use- and disease severity-dependent manner.
authors.affiliation: Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037 Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037.
authors.forename: David S
authors.initials: DS
authors.lastname: Gokhin
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Tierney
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Sui
authors.forename: Alessandra
authors.initials: A
authors.lastname: Sacco
authors.forename: Velia M
authors.initials: VM
authors.lastname: Fowler
mesh_terms: D008841:Actin Cytoskeleton / Q000235:genetics / Q000378:metabolism; D000185:Actinin / Q000235:genetics / Q000378:metabolism; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002154:Calpain / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D051379:Mice; D018101:Mice, Inbred mdx; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D017434:Protein Structure, Tertiary; D059748:Proteolysis; D051348:Tropomodulin / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D009124:Muscle Proteins; C495831:Tmod1 protein, mouse; C495837:Tmod3 protein, mouse; C495840:Tmod4 protein, mouse; D051348:Tropomodulin; C035620:nebulin; D000185:Actinin; D002154:Calpain; C449000:m-calpain