Post-translational modification of alpha-tubulin by acetylation and detyrosination in NB2a/d1 neuroblastoma cells.
Western blot analyses of total assembled microtubule fractions from NB2a/d1 neuroblastoma cells demonstrated that these cells are capable of post-translationally modifying alpha-tubulin by acetylation and detyrosination. Immunocytochemical analyses of NB2a/d1 cells differentiated with dbcAMP which had been processed under microtubule-stabilizing conditions demonstrated that all forms of alpha-tubulin were present throughout perikarya and neurites. By contrast, extraction of cells with Triton X-100 revealed a regional concentration of acetylated and detyrosinated alpha-tubulin subunits within axonal neurites, detectable in some cells after 3 days of differentiation and in nearly all cells after 7 days. Resistance of neurites to retraction following colchicine-treatment developed at a similar rate; furthermore, colchicine-resistant neurites contained intense acetylated alpha-tubulin immunoreactivity. We conclude that NB2a/d1 cells are capable of acetylating and detyrosinating alpha-tubulin subunits and that selective post-translational modification of alpha-tubulin subunits may be related to neuritic maturation.
Shea
T B
TB
Ralph Lowell Laboratories, McLean Hospital, Belmont, MA 02178.
Beermann
M L
ML
Nixon
R A
RA
D000107:Acetylation; D000818:Animals; D015153:Blotting, Western; D051379:Mice; D009447:Neuroblastoma; D011499:Protein Processing, Post-Translational; D014404:Tubulin / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
D014404:Tubulin; D014443:Tyrosine