Dysferlin interacts with tubulin and microtubules in mouse skeletal muscle.
Dysferlin is a type II transmembrane protein implicated in surface membrane repair in muscle. Mutations in dysferlin lead to limb girdle muscular dystrophy 2B, Miyoshi Myopathy and distal anterior compartment myopathy. Dysferlin's mode of action is not well understood and only a few protein binding partners have thus far been identified. Using affinity purification followed by liquid chromatography/mass spectrometry, we identified alpha-tubulin as a novel binding partner for dysferlin. The association between dysferlin and alpha-tubulin, as well as between dysferlin and microtubules, was confirmed in vitro by glutathione S-transferase pulldown and microtubule binding assays. These interactions were confirmed in vivo by co-immunoprecipitation. Confocal microscopy revealed that dysferlin and alpha-tubulin co-localized in the perinuclear region and in vesicular structures in myoblasts, and along thin longitudinal structures reminiscent of microtubules in myotubes. We mapped dysferlin's alpha-tubulin-binding region to its C2A and C2B domains. Modulation of calcium levels did not affect dysferlin binding to alpha-tubulin, suggesting that this interaction is calcium-independent. Our studies identified a new binding partner for dysferlin and suggest a role for microtubules in dysferlin trafficking to the sarcolemma.
Azakir
Bilal A
BA
Neuromuscular Research Group, Montreal Neurological Institute and Hospital, McGill University, Montreal, Quebec, Canada.
Di Fulvio
Sabrina
S
Therrien
Christian
C
Sinnreich
Michael
M
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D000073939:Dysferlin; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism*; D032446:Myoblasts / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D014404:Tubulin / Q000378:metabolism*
C497505:Dysf protein, mouse; D000073939:Dysferlin; D008565:Membrane Proteins; D014404:Tubulin; D002118:Calcium
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