Alteration of the C-terminal amino acid of tubulin specifically inhibits myogenic differentiation.
Detyrosination is an evolutionarily conserved post-translational modification of microtubule polymers that is known to be enhanced during early morphological differentiation of cultured myogenic cells (Gundersen, G. G., Khawaja, S., and Bulinski, J. C. (1989) J. Cell Biol. 109, 2275-2288). We proposed that altering the C terminus of alpha-tubulin by detyrosination plays a role in morphological differentiation. To test our hypothesis, we treated L6 myoblasts with 3-nitrotyrosine (Eiserich, J. P., Estevez, A. G., Bamberg, T. V., Ye, Y. Z., Chumley, P. H., Beckman, J. S., and Freeman, B. A. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 6365-6375), a nontoxic inhibitor that resulted in high level inhibition of microtubule detyrosination and low level incorporation of nitrotyrosine into microtubules. Even though microtubule stabilization or modification by acetylation still occurred normally, morphological differentiation was blocked; myoblasts neither elongated significantly nor fused. Nitrotyrosine treatment prevented synthesis or activation of markers of myogenic differentiation, including muscle-specific myosin, alpha-actin, integrin alpha(7), and myogenin. Consistent with this, myoblast integrin beta(1A) remained highly expressed. In contrast, the increase in beta-catenin level characteristic of early myogenesis was unaffected by treatment. These results show that the identity of the C-terminal residue of alpha-tubulin modulates microtubule activity, possibly because binding to or signaling from modified microtubules is required for the myogenic program.
Chang
Winston
W
Department of Biological Sciences, College of Arts & Sciences, Columbia University, New York, NY 10027-2450, USA.
Webster
Daniel R
DR
Salam
Ambar A
AA
Gruber
Dorota
D
Prasad
Aparna
A
Eiserich
Jason P
JP
Bulinski
J Chloë
JC
D000818:Animals; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D002454:Cell Differentiation; D002478:Cells, Cultured; D006863:Hydrogen-Ion Concentration; D009024:Morphogenesis / Q000187:drug effects; D024510:Muscle Development / Q000502:physiology*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D051381:Rats; D014404:Tubulin / Q000737:chemistry* / Q000502:physiology; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
D015815:Cell Adhesion Molecules; D014404:Tubulin; C002744:3-nitrotyrosine; D014443:Tyrosine; D002268:Carboxypeptidases; C023212:tyrosyltubulin carboxypeptidase