The clathrin-dependent spindle proteome

Sushma R. Rao, Neftali Flores-Rodriguez, Scott L. Page, Chin Wong, Phillip J. Robinson, Megan Chircop

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The mitotic spindle is required for chromosome congression and subsequent equal segregation of sister chromatids. These processes involve a complex network of signaling molecules located at the spindle. The endocytic protein, clathrin, has a "moonlighting" role during mitosis, whereby it stabilizes the mitotic spindle. The signaling pathways that clathrin participates in to achieve mitotic spindle stability are unknown. Here, we assessed the mitotic spindle proteome and phosphoproteome in clathrindepleted cells using quantitative MS/MS (data are available via ProteomeXchange with identifier PXD001603). We report a spindle proteome that consists of 3046 proteins and a spindle phosphoproteome consisting of 5157 phosphosites in 1641 phosphoproteins. Of these, 2908 (95.4%) proteins and 1636 (99.7%) phosphoproteins are known or predicted spindle-associated proteins. Clathrin-depletion from spindles resulted in dysregulation of 121 proteins and perturbed signaling to 47 phosphosites. The majority of these proteins increased in mitotic spindle abundance and six of these were validated by immunofluorescence microscopy. Functional pathway analysis confirmed the reported role of clathrin in mitotic spindle stabilization for chromosome alignment and highlighted possible new mechanisms of clathrin action. The data also revealed a novel second mitotic role for clathrin in bipolar spindle formation.

Original languageEnglish
Pages (from-to)2537-2553
Number of pages17
JournalMolecular and Cellular Proteomics
Volume15
Issue number8
DOIs
Publication statusPublished - Aug 2016
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

Cite this

Rao, S. R., Flores-Rodriguez, N., Page, S. L., Wong, C., Robinson, P. J., & Chircop, M. (2016). The clathrin-dependent spindle proteome. Molecular and Cellular Proteomics, 15(8), 2537-2553. https://doi.org/10.1074/mcp.M115.054809