The nuclear localization of γ-tubulin is regulated by SadB-mediated phosphorylation

J Biol Chem. 2014 Aug 1;289(31):21360-73. doi: 10.1074/jbc.M114.562389. Epub 2014 Jun 18.

Abstract

γ-Tubulin is an important cell division regulator that arranges microtubule assembly and mitotic spindle formation. Cytosolic γ-tubulin nucleates α- and β-tubulin in a growing microtubule by forming the ring-shaped protein complex γTuRC. Nuclear γ-tubulin also regulates S-phase progression by moderating the activities of E2 promoter-binding factors. The mechanism that regulates localization of γ-tubulin is currently unknown. Here, we demonstrate that the human Ser/Thr kinase SadB short localizes to chromatin and centrosomes. We found that SadB-mediated phosphorylation of γ-tubulin on Ser(385) formed chromatin-associated γ-tubulin complexes that moderate gene expression. In this way, the C-terminal region of γ-tubulin regulates S-phase progression. In addition, chromatin levels of γ-tubulin were decreased by the reduction of SadB levels or expression of a non-phosphorylatable Ala(385)-γ-tubulin but were enhanced by expression of SadB, wild-type γ-tubulin, or a phosphomimetic Asp(385)-γ-tubulin mutant. Our results demonstrate that SadB kinases regulate the cellular localization of γ-tubulin and thereby control S-phase progression.

Keywords: Cell Cycle; Centrosome; Nucleus; Phosphorylation; Tubulin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line, Tumor
  • Cell Nucleus / metabolism*
  • DNA Primers
  • Gene Expression / physiology
  • Humans
  • Mice
  • Molecular Sequence Data
  • NIH 3T3 Cells
  • Phosphorylation
  • Polymerase Chain Reaction
  • Protein Kinases / chemistry
  • Protein Kinases / metabolism*
  • Sequence Homology, Amino Acid
  • Tubulin / metabolism*
  • Tubulin / physiology

Substances

  • DNA Primers
  • Tubulin
  • Protein Kinases