Mdm muscular dystrophy: interactions with calpain 3 and a novel functional role for titin's N2A domain.

Human tibial muscular dystrophy and limb-girdle muscular dystrophy 2J are caused by mutations in the giant sarcomeric protein titin (TTN) adjacent to a binding site for the muscle-specific protease calpain 3 (CAPN3). Muscular dystrophy with myositis (mdm) is a recessive mouse mutation with severe and progressive muscular degeneration caused by a deletion in the N2A domain of titin (TTN-N2ADelta83), disrupting a putative binding site for CAPN3. To determine whether the muscular dystrophy in mutant mdm mice is caused by misregulation of CAPN3 activity, genetic crosses with CAPN3 overexpressing transgenic (C3Tg) and CAPN3 knockout (C3KO) mice were generated. Here, we report that overexpression of CAPN3 exacerbates the mdm disease, leading to a shorter life span and more severe muscular dystrophy. However, in a direct genetic test of CAPN3's role as a mediator of mdm pathology, C3KO;mdm double mutant mice showed no change in the progression or severity of disease indicating that aberrant CAPN3 activity is not a primary mechanism in this disease. To determine whether we could detect a functional deficit in titin in a non-disease state, we examined the treadmill locomotion of heterozygous +/mdm mice and detected a significant increase in stride time with a concomitant increase in stance time. Interestingly, these altered gait parameters were completely corrected by CAPN3 overexpression in transgenic C3Tg;+/mdm mice, supporting a CAPN3-dependent role for the N2A domain of TTN in the dynamics of muscle contraction.

[1]  W. Frankel,et al.  Gait analysis detects early changes in transgenic SOD1(G93A) mice , 2005, Muscle & nerve.

[2]  Thomas Sejersen,et al.  The Kinase Domain of Titin Controls Muscle Gene Expression and Protein Turnover , 2005, Science.

[3]  E. Kudryashova,et al.  Null mutation of calpain 3 (p94) in mice causes abnormal sarcomere formation in vivo and in vitro. , 2004, Human molecular genetics.

[4]  C. Gregorio,et al.  Muscle-specific RING finger-2 (MURF-2) is important for microtubule, intermediate filament and sarcomeric M-line maintenance in striated muscle development , 2004, Journal of Cell Science.

[5]  Giorgio Valle,et al.  The Ankrd2 protein, a link between the sarcomere and the nucleus in skeletal muscle. , 2004, Journal of molecular biology.

[6]  Yiming Wu,et al.  Induction and myofibrillar targeting of CARP, and suppression of the Nkx2.5 pathway in the MDM mouse with impaired titin-based signaling. , 2004, Journal of molecular biology.

[7]  Christian C Witt,et al.  The muscle ankyrin repeat proteins: CARP, ankrd2/Arpp and DARP as a family of titin filament-based stress response molecules. , 2003, Journal of molecular biology.

[8]  D. E. Goll,et al.  The calpain system. , 2003, Physiological reviews.

[9]  Marion L Greaser,et al.  Vertical agarose gel electrophoresis and electroblotting of high‐molecular‐weight proteins , 2003, Electrophoresis.

[10]  K. Campbell,et al.  Dystrophin-Glycoprotein Complex: Post-translational Processing and Dystroglycan Function* 210 , 2003, The Journal of Biological Chemistry.

[11]  I. Nishino,et al.  Localization of calpain 3 in human skeletal muscle and its alteration in limb-girdle muscular dystrophy 2A muscle. , 2003, Journal of biochemistry.

[12]  M. Matsuo,et al.  Molecular Identification and Characterization of a Novel Nuclear Protein Whose Expression Is Up-regulated in Insulin-resistant Animals* , 2003, The Journal of Biological Chemistry.

[13]  J. Beckmann,et al.  The 105th ENMC sponsored workshop: pathogenesis in the non-sarcoglycan limb-girdle muscular dystrophies, Naarden, April 12–14, 2002 , 2003, Neuromuscular Disorders.

[14]  Ichizo Nishino,et al.  Muscular dystrophies , 2002, Current opinion in neurology.

[15]  D. Skuk,et al.  Experimental and therapeutic approaches to muscular dystrophies , 2002, Current opinion in neurology.

[16]  Leena Peltonen,et al.  Tibial muscular dystrophy is a titinopathy caused by mutations in TTN, the gene encoding the giant skeletal-muscle protein titin. , 2002, American journal of human genetics.

[17]  Paul Keim,et al.  Do muscles function as adaptable locomotor springs? , 2002, The Journal of experimental biology.

[18]  J. Beckmann,et al.  Stable expression of calpain 3 from a muscle transgene in vivo: Immature muscle in transgenic mice suggests a role for calpain 3 in muscle maturation , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[19]  C. Gregorio,et al.  Muscle-specific RING finger-1 interacts with titin to regulate sarcomeric M-line and thick filament structure and may have nuclear functions via its interaction with glucocorticoid modulatory element binding protein-1 , 2002, The Journal of cell biology.

[20]  Wayne N Frankel,et al.  The muscular dystrophy with myositis (mdm) mouse mutation disrupts a skeletal muscle-specific domain of titin. , 2002, Genomics.

[21]  Dietmar Labeit,et al.  The Complete Gene Sequence of Titin, Expression of an Unusual ≈700-kDa Titin Isoform, and Its Interaction With Obscurin Identify a Novel Z-Line to I-Band Linking System , 2001 .

[22]  M. Bang,et al.  Specific interaction of the potassium channel beta-subunit minK with the sarcomeric protein T-cap suggests a T-tubule-myofibril linking system. , 2001, Journal of molecular biology.

[23]  L. Peltonen,et al.  Secondary calpain3 deficiency in 2q-linked muscular dystrophy , 2001, Neurology.

[24]  K. Pelin,et al.  Identification of muscle specific ring finger proteins as potential regulators of the titin kinase domain. , 2001, Journal of molecular biology.

[25]  J. Beckmann,et al.  Loss of Calpain 3 Proteolytic Activity Leads to Muscular Dystrophy and to Apoptosis-Associated Iκbα/Nuclear Factor κb Pathway Perturbation in Mice , 2000, The Journal of cell biology.

[26]  J. Gilbert,et al.  Myotilin is mutated in limb girdle muscular dystrophy 1A. , 2000, Human molecular genetics.

[27]  R. Nagai,et al.  Cardiac ankyrin repeat protein is a novel marker of cardiac hypertrophy: role of M-CAT element within the promoter. , 2000, Hypertension.

[28]  T. Kemp,et al.  Identification of Ankrd2, a novel skeletal muscle gene coding for a stretch-responsive ankyrin-repeat protein. , 2000, Genomics.

[29]  T Centner,et al.  Series of exon-skipping events in the elastic spring region of titin as the structural basis for myofibrillar elastic diversity. , 2000, Circulation research.

[30]  K. Suzuki,et al.  Myopathy phenotype of transgenic mice expressing active site-mutated inactive p94 skeletal muscle-specific calpain, the gene product responsible for limb girdle muscular dystrophy type 2A. , 2000, Human molecular genetics.

[31]  G. Valle,et al.  Limb-girdle muscular dystrophy type 2G is caused by mutations in the gene encoding the sarcomeric protein telethonin , 2000, Nature Genetics.

[32]  J. Beckmann,et al.  Expression and Functional Characteristics of Calpain 3 Isoforms Generated through Tissue-Specific Transcriptional and Posttranscriptional Events , 1999, Molecular and Cellular Biology.

[33]  Pico Caroni,et al.  Accumulation of Muscle Ankyrin Repeat Protein Transcript Reveals Local Activation of Primary Myotube Endcompartments during Muscle Morphogenesis , 1997, The Journal of cell biology.

[34]  K. Suzuki,et al.  Muscle-specific calpain, p94, interacts with the extreme C-terminal region of connectin, a unique region flanked by two immunoglobulin C2 motifs. , 1997, Archives of biochemistry and biophysics.

[35]  H. Granzier,et al.  Titin develops restoring force in rat cardiac myocytes. , 1996, Circulation research.

[36]  G. Ettema.  Elastic and length-force characteristics of the gastrocnemius of the hopping mouse (Notomys alexis) and the rat (Rattus norvegicus). , 1996, The Journal of experimental biology.

[37]  H. Jockusch,et al.  Overshooting production of satellite cells in murine skeletal muscle affected by the mutation ”muscular dystrophy with myositis” (mdm, Chr 2) , 1996, Cell and Tissue Research.

[38]  H. Sorimachi,et al.  Muscle-specific Calpain, p94, Responsible for Limb Girdle Muscular Dystrophy Type 2A, Associates with Connectin through IS2, a p94-specific Sequence (*) , 1995, The Journal of Biological Chemistry.

[39]  Siegfried Labeit,et al.  Titins: Giant Proteins in Charge of Muscle Ultrastructure and Elasticity , 1995, Science.

[40]  A. Nishikawa,et al.  Visualization of dystrophic muscle fibers in mdx mouse by vital staining with Evans blue: evidence of apoptosis in dystrophin-deficient muscle. , 1995, Journal of biochemistry.

[41]  Isabelle Richard,et al.  Mutations in the proteolytic enzyme calpain 3 cause limb-girdle muscular dystrophy type 2A , 1995, Cell.

[42]  H. Kawasaki,et al.  Muscle-specific calpain, p94, is degraded by autolysis immediately after translation, resulting in disappearance from muscle. , 1993, The Journal of biological chemistry.

[43]  K. Wang,et al.  Regulation of skeletal muscle stiffness and elasticity by titin isoforms: a test of the segmental extension model of resting tension. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[44]  L. Edström,et al.  Myopathy with respiratory failure and typical myofibrillar lesions , 1990, Journal of the Neurological Sciences.

[45]  K. Weber,et al.  The organization of titin filaments in the half-sarcomere revealed by monoclonal antibodies in immunoelectron microscopy: a map of ten nonrepetitive epitopes starting at the Z line extends close to the M line , 1988, The Journal of cell biology.

[46]  A. Hattori,et al.  Calcium binding to an elastic portion of connectin/titin filaments , 2004, Journal of Muscle Research & Cell Motility.

[47]  T Centner,et al.  The complete gene sequence of titin, expression of an unusual approximately 700-kDa titin isoform, and its interaction with obscurin identify a novel Z-line to I-band linking system. , 2001, Circulation research.

[48]  K. Suzuki,et al.  Skeletal muscle-specific calpain, p94, and connectin/titin: their physiological functions and relationship to limb-girdle muscular dystrophy type 2A. , 2000, Advances in Experimental Medicine and Biology.

[49]  J. Trinick.  Titin as a scaffold and spring. Cytoskeleton. , 1996, Current biology : CB.

[50]  A. Hattori,et al.  Detection of giant myofibrillar proteins connectin and nebulin by electrophoresis in 2% polyacrylamide slab gels strengthened with agarose. , 1995, Analytical biochemistry.