A centrosomal protein FOR20 regulates microtubule assembly dynamics and plays a role in cell migration

Biochem J. 2017 Aug 10;474(16):2841-2859. doi: 10.1042/BCJ20170303.

Abstract

Here, we report that a centrosomal protein FOR20 [FOP (FGFR1 (fibroblast growth factor receptor 1) oncogene protein)-like protein of molecular mass of 20 kDa; also named as C16orf63, FLJ31153 or PHSECRG2] can regulate the assembly and stability of microtubules. Both FOR20 IgG antibody and GST (glutathione S-transferase)-tagged FOR20 could precipitate tubulin from the HeLa cell extract, indicating a possible interaction between FOR20 and tubulin. FOR20 was also detected in goat brain tissue extract and it cycled with microtubule-associated proteins. Furthermore, FOR20 bound to purified tubulin and inhibited the assembly of tubulin in vitro. The overexpression of FOR20 depolymerized interphase microtubules and the depletion of FOR20 prevented nocodazole-induced depolymerization of microtubules in HeLa cells. In addition, the depletion of FOR20 suppressed the dynamics of individual microtubules in live HeLa cells. FOR20-depleted MDA-MB-231 cells displayed zigzag motion and migrated at a slower rate than the control cells, indicating that FOR20 plays a role in directed cell migration. The results suggested that the centrosomal protein FOR20 is a new member of the microtubule-associated protein family and that it regulates the assembly and dynamics of microtubules.

Keywords: cell motility; centrosome; microtubule dynamics; microtubule-associated proteins.

MeSH terms

  • Animals
  • Brain Chemistry
  • Cell Line, Tumor
  • Cell Movement
  • Cilia / metabolism*
  • Cilia / ultrastructure
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Epithelial Cells / ultrastructure
  • Focal Adhesions / drug effects
  • Focal Adhesions / metabolism
  • Focal Adhesions / ultrastructure
  • Gene Expression
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Goats
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Mice
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism*
  • Microtubules / ultrastructure
  • NIH 3T3 Cells
  • Nocodazole / pharmacology
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / metabolism
  • Proteins / genetics*
  • Proteins / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Time-Lapse Imaging
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Tubulin / genetics*
  • Tubulin / metabolism

Substances

  • CEP20 protein, human
  • CHTOP protein, human
  • MAPRE1 protein, human
  • Microtubule-Associated Proteins
  • Nuclear Proteins
  • Proteins
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Tubulin
  • Green Fluorescent Proteins
  • Glutathione Transferase
  • Nocodazole