title: Disease-associated mutations in TUBA1A result in a spectrum of defects in the tubulin folding and heterodimer assembly pathway.
abstract: Malformations of cortical development are characteristic of a plethora of diseases that includes polymicrogyria, periventricular and subcortical heterotopia and lissencephaly. Mutations in TUBA1A and TUBB2B, each a member of the multigene families that encode alpha- and beta-tubulins, have recently been implicated in these diseases. Here we examine the defects that result from nine disease-causing mutations (I188L, I238V, P263T, L286F, V303G, L397P, R402C, 402H, S419L) in TUBA1A. We show that the expression of all the mutant proteins in vitro results in the generation of tubulin heterodimers in varying yield and that these can co-polymerize with microtubules in vitro. We identify several kinds of defects that result from these mutations. Among these are various defects in the chaperone-dependent pathway leading to de novo tubulin heterodimer formation. These include a defective interaction with the chaperone prefoldin, a reduced efficiency in the generation of productive folding intermediates as a result of inefficient interaction with the cytosolic chaperonin, CCT, and, in several cases, a failure to stably interact with TBCB, one of five tubulin-specific chaperones that act downstream of CCT in the tubulin heterodimer assembly pathway. Other defects include structural instability in vitro, diminished stability in vivo, a compromised ability to co-assemble with microtubules in vivo and a suppression of microtubule growth rate in the neurites (but not the soma) of cultured neurons. Our data are consistent with the notion that some mutations in TUBA1A result in tubulin deficit, whereas others reflect compromised interactions with one or more MAPs that are essential to proper neuronal migration.
authors.affiliation: Department of Biochemistry, NYU Langone Medical Center, New York, NY 10016, USA.
authors.forename: Guoling
authors.initials: G
authors.lastname: Tian
authors.forename: Xavier H
authors.initials: XH
authors.lastname: Jaglin
authors.forename: David A
authors.initials: DA
authors.lastname: Keays
authors.forename: Fiona
authors.initials: F
authors.lastname: Francis
authors.forename: Jamel
authors.initials: J
authors.lastname: Chelly
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Cowan
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D019281:Dimerization; D006801:Humans; D054220:Malformations of Cortical Development / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008968:Molecular Conformation; D009154:Mutation; D020125:Mutation, Missense; D017510:Protein Folding; D055550:Protein Stability; D014404:Tubulin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: C533522:TUBA1A protein, human; D014404:Tubulin