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

• Mirko Theis ^[2] ; Mikolaj Slabicki ^[2] ; Magno Junqueira ^[2] ; Maciej Paszkowski-Rogacz ^[2] ; Jana Sontheimer ^[1] ; Ralf Kittler ^[2] ; Anne-Kristine Heninger ^[2] ; Timo Glatter ^[3] ; Kristi Kruusmaa ^[2] ; Ina Poser ^[2] ; Anthony A Hyman ^[2] ; M Teresa Pisabarro ^[1] ; Matthias Gstaiger ^[3] ; Rudolf Aebersold ^[3] ; Andrej Shevchenko ^[2] ; Frank Buchholz ^[2]
  1. [1] Dresden University of Technology

     Dresden University of Technology

     Kreisfreie Stadt Dresden, Alemania

     
  2. [2] Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany
  3. [3] Institute for Molecular Systems Biology, ETH, Zürich, Switzerland

  Mostrar afiliaciones +
• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 28, Nº. 10, 2009, págs. 1453-1465
• Idioma: inglés
• DOI: 10.1038/emboj.2009.114
• Enlaces
  • Texto completo
• Resumen

  • 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.

• Referencias bibliográficas
  • (1996). Genetic characterization of a mammalian protein–protein interaction domain by using a yeast reverse two-hybrid system. Proc Natl Acad...
  • (1996). Threading thrills and threats. Structure. 4. 15-19
  • (1999). Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation. J Biol Chem. 274. 25543-25549
  • (2000). DNA cloning by homologous recombination in. Nat Biotechnol. 18. 1314-1317
  • (2000). Two distinct pathways remove mammalian cohesion from chromosome arms in prophase and from centromeres in anaphase. Cell. 103. 399-410
  • (2001). Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol. 2. 21-32
  • (2001). Techniques: recombinogenic engineering—new options for cloning and manipulating DNA. Trends Biochem Sci. 26. 325-331
  • (2002). Segregating sister genomes: the molecular biology of chromosome separation. Science. 297. 559-565
  • (2002). Short RNA duplexes produced by hydrolysis with. Proc Natl Acad Sci USA. 99. 9942-9947
  • (2003). ELM server: a new resource for investigating short functional sites in modular eukaryotic proteins. Nucleic Acids Res. 31. 3625-3630
  • (2003). Fold recognition methods. Methods Biochem Anal. 44. 525-546
  • (2003). Global analysis of protein localization in budding yeast. Nature. 425. 686-691
  • (2003). Okadaic acid, an inhibitor of protein phosphatase 1 and 2A, induces premature separation of sister chromatids during meiosis I and...
  • (2004). An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Nature. 432. 1036-1040
  • (2004). Oncology studies using siRNA libraries: the dawn of RNAi-based genomics. Oncogene. 23. 8384-8391
  • (2004). Robust phosphoproteomic profiling of tyrosine phosphorylation sites from human T cells using immobilized metal affinity chromatography...
  • (2005). A combined approach for the localization and tandem affinity purification of protein complexes from metazoans. Sci STKE. 2005. 1
  • (2005). Immunoaffinity profiling of tyrosine phosphorylation in cancer cells. Nat Biotechnol. 23. 94-101
  • (2005). Production of endoribonuclease-prepared short interfering RNAs for gene silencing in mammalian cells. Nat Methods. 2. 779-784
  • (2005). RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells. Proc Natl Acad Sci USA....
  • (2005). Shugoshin: guardian spirit at the centromere. Curr Opin Cell Biol. 17. 590-595
  • (2006). An architectural map of the anaphase-promoting complex. Genes Dev. 20. 449-460
  • (2006). Global landscape of protein complexes in the yeast. Nature. 440. 637-643
  • (2006). High-throughput fluorescence microscopy for systems biology. Nat Rev Mol Cell Biol. 7. 690-696
  • (2006). Phosphoproteome analysis of the human mitotic spindle. Proc Natl Acad Sci USA. 103. 5391-5396
  • (2006). Protein phosphatase 2A protects centromeric sister chromatid cohesion during meiosis I. Nature. 441. 53-61
  • (2006). Proteome survey reveals modularity of the yeast cell machinery. Nature. 440. 631-636
  • (2006). Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc Natl Acad Sci USA. 103. 10660-10665
  • (2006). Structural analysis of the interaction between the SNARE Tlg1 and Vps51. Traffic. 7. 182-190
  • (2006). The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat Rev Mol Cell Biol. 7. 644-656
  • (2006). Timely anaphase onset requires a novel spindle and kinetochore complex comprising Ska1 and Ska2. EMBO J. 25. 5504-5515
  • (2007). Finishing mitosis, one step at a time. Nat Rev Mol Cell Biol. 8. 894-903
  • (2007). Genome-scale RNAi profiling of cell division in human tissue culture cells. Nat Cell Biol. 9. 1401-1412
  • (2007). Genome-wide resources of endoribonuclease-prepared short interfering RNAs for specific loss-of-function studies. Nat Methods. 4. 337-344
  • (2007). Structural analysis of Bub3 interactions in the mitotic spindle checkpoint. Proc Natl Acad Sci USA. 104. 1201-1206
  • (2007). The complete removal of cohesin from chromosome arms depends on separase. J Cell Sci. 120. 4188-4196
  • (2007). The spindle-assembly checkpoint in space and time. Nat Rev Mol Cell Biol. 8. 379-393
  • (2007). siRNA screen of the human signaling proteome identifies the PtdIns(3,4,5)P3-mTOR signaling pathway as a primary regulator of transferrin...
  • (2008). A protein domain-based interactome network for. Cell. 134. 534-545
  • (2008). An RNAi screen of chromatin proteins identifies Tip60-p400 as a regulator of embryonic stem cell identity. Cell. 134. 162-174
  • (2008). BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. Nat Methods. 5. 409-415
  • (2008). Combining protein-based IMAC, peptide-based IMAC, and MudPIT for efficient phosphoproteomic analysis. J Proteome Res. 7. 1346-1351
  • (2008). Discovery of candidate KEN-box motifs using cell cycle keyword enrichment combined with native disorder prediction and motif conservation....
  • (2008). Fluorescence proteins, live-cell imaging, and mechanobiology: seeing is believing. Annu Rev Biomed Eng. 10. 1-38
  • (2008). Heterochromatin links to centromeric protection by recruiting shugoshin. Nature. 455. 251-255
  • (2008). Systems biology of mammalian cell division. Cell Cycle. 7. 2123-2128
  • (2008). The Jpred 3 secondary structure prediction server. Nucleic Acids Res. 36. 197-201
  • (2008). The RosettaDock server for local protein–protein docking. Nucleic Acids Res. 36. 233-238
  • (2008). The spindle assembly checkpoint. Curr Biol. 18. 591-595
  • (2008). Whole genome functional analysis identifies novel components required for mitotic spindle integrity in human cells. Genome Biol. 9....
  • (2008). sSgo1, a major splice variant of Sgo1, functions in centriole cohesion where it is regulated by Plk1. Dev Cell. 14. 331-341
  • (2009). An integrated workflow for charting the human interaction proteome: insights into the PP2A system. Mol Syst Biol. 5. 237