Microtubule-associated protein 4 (MAP4) regulates assembly, protomer-polymer partitioning and synthesis of tubulin in cultured cells.
We depleted MAP4, a ubiquitously expressed microtubule (MT)-associated protein previously shown to be capable of stabilizing MTs, from HeLa cells by stably expressing antisense RNA. These HeLa-AS cells, in which the MAP4 level was decreased to 33% of the wild-type level, displayed decreased content of total tubulin (65% of the wild-type level). The partitioning of cellular tubulin into protomer and polymer was altered in HeLa-AS cells: polymeric tubulin was decreased to 46% of the level in control cells, while protomeric tubulin was increased to 226% of the level in control cells. Tubulin protein synthesis was decreased, consistent with the tubulin autoregulation model, which proposes that tubulin protomer inhibits its own synthesis. Following release from drug-induced depolymerization, MTs in HeLa-AS cells reformed more slowly, and showed an increased focus on the centrosome, as compared to control cells. HeLa-AS cells also appeared to be less bipolar in shape and flatter than control cells. Our data suggest that MAP4 regulates assembly level of MTs and, perhaps through this mechanism, is involved in controlling spreading and shape of cells.
Nguyen
H L
HL
Departments of Pathology and Anatomy and Cell Biology, Columbia University, College of Physicians and Surgeons, BB1213-630 W. 168th St, New York, NY 10032-3702, USA. jcb4@columbia.edu
Gruber
D
D
Bulinski
J C
JC
D001704:Biopolymers; D006367:HeLa Cells; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014404:Tubulin / Q000096:biosynthesis*
D001704:Biopolymers; D008869:Microtubule-Associated Proteins; D014404:Tubulin