Comparative profiling identifies C13orf3 as a component of the Ska complex required for mammalian cell division

Mirko Theis, Mikolaj Slabicki, Magno Junqueira, MacIej Paszkowski-Rogacz, Jana Sontheimer, Ralf Kittler, Anne Kristine Heninger, Timo Glatter, Kristi Kruusmaa, Ina Poser, Anthony A. Hyman, M. Teresa Pisabarro, Matthias Gstaiger, Rudolf Aebersold, Andrej Shevchenko, Frank Buchholz

Research output: Contribution to journalArticle

67 Scopus citations

Abstract

Proliferation of mammalian cells requires the coordinated function of many proteins to accurately divide a cell into two daughter cells. Several RNAi screens have identified previously uncharacterised genes that are implicated in mammalian cell division. The molecular function for these genes needs to be investigated to place them into pathways. Phenotypic profiling is a useful method to assign putative functions to uncharacterised genes. Here, we show that the analysis of protein localisation is useful to refine a phenotypic profile. We show the utility of this approach by defining a function of the previously uncharacterised gene C13orf3 during cell division. C13orf3 localises to centrosomes, the mitotic spindle, kinetochores, spindle midzone, and the cleavage furrow during cell division and is specifically phosphorylated during mitosis. Furthermore, C13orf3 is required for centrosome integrity and anaphase onset. Depletion by RNAi leads to mitotic arrest in metaphase with an activation of the spindle assembly checkpoint and loss of sister chromatid cohesion. Proteomic analyses identify C13orf3 (Ska3) as a new component of the Ska complex and show a direct interaction with a regulatory subunit of the protein phosphatase PP2A. All together, these data identify C13orf3 as an important factor for metaphase to anaphase progression and highlight the potential of combined RNAi screening and protein localisation analyses.

Original languageEnglish (US)
Pages (from-to)1453-1465
Number of pages13
JournalEMBO Journal
Volume28
Issue number10
DOIs
StatePublished - May 20 2009

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Keywords

  • EsiRNA
  • Kinetochore
  • Shugoshin
  • Ska
  • Spindle checkpoint

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Theis, M., Slabicki, M., Junqueira, M., Paszkowski-Rogacz, M., Sontheimer, J., Kittler, R., Heninger, A. K., Glatter, T., Kruusmaa, K., Poser, I., Hyman, A. A., Pisabarro, M. T., Gstaiger, M., Aebersold, R., Shevchenko, A., & Buchholz, F. (2009). Comparative profiling identifies C13orf3 as a component of the Ska complex required for mammalian cell division. EMBO Journal, 28(10), 1453-1465. https://doi.org/10.1038/emboj.2009.114