title: The nebulin SH3 domain is dispensable for normal skeletal muscle structure but is required for effective active load bearing in mouse.
abstract: Nemaline myopathy (NM) is a congenital myopathy with an estimated incidence of 150,000 live births. It is caused by mutations in thin filament components, including nebulin, which accounts for about 50% of the cases. The identification of NM cases with nonsense mutations resulting in loss of the extreme C-terminal SH3 domain of nebulin suggests an important role of the nebulin SH3 domain, which is further supported by the recent demonstration of its role in IGF-1-induced sarcomeric actin filament formation through targeting of N-WASP to the Z-line. To provide further insights into the functional significance of the nebulin SH3 domain in the Z-disk and to understand the mechanisms by which truncations of nebulin lead to NM, we took two approaches: (1) an affinity-based proteomic screening to identify novel interaction partners of the nebulin SH3 domain; and (2) generation and characterization of a novel knockin mouse model with a premature stop codon in the nebulin gene, eliminating its C-terminal SH3 domain (NebΔSH3 mouse). Surprisingly, detailed analyses of NebΔSH3 mice revealed no structural or histological skeletal muscle abnormalities and no changes in gene expression or localization of interaction partners of the nebulin SH3 domain, including myopalladin, palladin, zyxin and N-WASP. Also, no significant effect on peak isometric stress production, passive tensile stress or Young's modulus was found. However, NebΔSH3 muscle displayed a slightly altered force-frequency relationship and was significantly more susceptible to eccentric contraction-induced injury, suggesting that the nebulin SH3 domain protects against eccentric contraction-induced injury and possibly plays a role in fine-tuning the excitation-contraction coupling mechanism.
authors.affiliation: Institute of Biomedical Technologies, National Research Council, 20090 Milan, Italy.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Yamamoto
authors.forename: Carmen
authors.initials: C
authors.lastname: Vitiello
authors.forename: Jianlin
authors.initials: J
authors.lastname: Zhang
authors.forename: David S
authors.initials: DS
authors.lastname: Gokhin
authors.forename: Alessandra
authors.initials: A
authors.lastname: Castaldi
authors.forename: Gerald
authors.initials: G
authors.lastname: Coulis
authors.forename: Fabio
authors.initials: F
authors.lastname: Piaser
authors.forename: Maria Carmela
authors.initials: MC
authors.lastname: Filomena
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Eggenhuizen
authors.forename: Paolo
authors.initials: P
authors.lastname: Kunderfranco
authors.forename: Serena
authors.initials: S
authors.lastname: Camerini
authors.forename: Kazunori
authors.initials: K
authors.lastname: Takano
authors.forename: Takeshi
authors.initials: T
authors.lastname: Endo
authors.forename: Marco
authors.initials: M
authors.lastname: Crescenzi
authors.forename: Pradeep K L
authors.initials: PK
authors.lastname: Luther
authors.forename: Richard L
authors.initials: RL
authors.lastname: Lieber
authors.forename: Ju
authors.initials: J
authors.lastname: Chen
authors.forename: Marie-Louise
authors.initials: ML
authors.lastname: Bang
mesh_terms: D000818:Animals; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D055119:Elastic Modulus / Q000502:physiology; D056966:Excitation Contraction Coupling / Q000502:physiology; D005260:Female; D015870:Gene Expression; D006801:Humans; D007537:Isometric Contraction / Q000502:physiology; D008297:Male; D051379:Mice; D009124:Muscle Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D017696:Myopathies, Nemaline / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013718:Tensile Strength / Q000502:physiology; D016474:Weight-Bearing / Q000502:physiology; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000235:genetics / Q000378:metabolism; D060589:Zyxin / Q000235:genetics / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D009124:Muscle Proteins; D010750:Phosphoproteins; C495772:Wasl protein, mouse; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; C498285:Zyx protein, mouse; D060589:Zyxin; C485873:palladin protein, mouse; C035620:nebulin
keywords: Nebulin; Nemaline myopathy; Sarcomere; Skeletal muscle; Z-line
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