Centrosome/Spindle

Watch Jeff Woodruff explain his project on Centrosome Assembly in his Two Minute Talk!

Kirkham, et al. SAS-4 is a C. elegans centriolar protein that controls centrosome size.
Cell 112, 575–587 (2003).

Control of Centrosome & Mitotic Spindle Assembly
We work on various aspects of centrosome maturation in C. elegans embryos.

Questions we address are:
• How is the pericentriolar matrix (PCM), an amorphous, micrometer-scale structure, assembled from thousands of nanometer-scale proteins in the cytoplasm? (See Woodruff et al 2015, Wueseke et al 2016 and its video abstract)
• What state do PCM components take before they form the PCM? (See Wueseke et al 2014)
• How does the centrosome disassemble during mitotic exit?

We take an interdisciplinary approach to addressing these questions, combining tools from genetics, live imaging, biophysics, bioinformatics and biochemistry.

Previous work on centrosomes in the lab focused on:
• How does the centrosome scale in size? (See Decker et al 2011, Goehring et al 2012)
• How does centrosome size influence the size of the mitotic spindle? (See Greenan et al 2010)
• What are the mechanisms underlying centriole assembly? (See Pelletier et al 2006)

Lab members working on centrosomes: Stephen Enos, Beatriz Ferreira Gomes, Andrea Zinke, and Jeff Woodruff

Recent Associated Publications (for more, visit our Publications Page)

    • Wueseke O, Zwicker D, Schwager A, Wong YL, Ogema K, Jülicher F, Hyman AA, Woodruff JB. Polo-like kinase phosphorylation determines C. elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation. Biology Open 2016 5:1431-1440. [FullText][VideoAbstract]
    • Woodruff JB, Wueseke O, Viscardi V, Mahamid J, Ochoa SD, Bunkenborg J, Widlund PO, Pozniakovsky A, Zanin E, Bahmanyar S,Zinke A, Hong SH, Decker M, Baumeister W, Andersen JS, Oegema K, Hyman AA. Centrosomes. Regulated assembly of a supramolecular centrosome scaffold in vitro.  Science. 2015 May 15;348(6236):808-12. [PubMed] (Please visit our Publications page for the referrer links to the free full text and PDF.)
      This work describes a novel in vitro system for studying regulated assembly of the pericentriolar matrix (PCM), revealing that networks of the protein SPD-5 can polymerize into interconnected, porous networks that specifically recruit PCM proteins.
    • Woodruff JB, Hyman AA. Method: In vitro analysis of pericentriolar material assembly.
      Methods Cell Biol. 2015;129:369-82. [PubMed] [PDF]
      A methods paper to further describe the technique behind the ‘Science’ paper above.
    • Wuekese O, Bunkenborg J, Hein MY, Zinke A, Viscardi V, Woodruff JB, Oegema K, Mann M, Andersen JS, Hyman AA. The C. elegans pericentriolar material components SPD-2 and SPD-5 are monomeric in the cytoplasm prior to incorporation into the PCM matrix. Mol Biol Cell. 2014 Aug 7. pii: mbc.E13-09-0514. [Epub ahead of print] [PubMed]
      PCM components interact only in the context of PCM assembly, rather than first forming smaller complexes in the cytoplasm.
    • Woodruff JB, Wueseke O, Hyman AA. Pericentriolar material structure and dynamics. Phil. Trans. R. Soc. B. 2014 July; 369, 20130459. [PDF]
      A review of the current knowledge about the PCM, as part of a special theme issue of Phil. Trans. R. Soc. B about centrosome biology, “The centrosome renaissance.
    • Zwicker D, Decker M, Jaensch S, Hyman AA, Jülicher F. Centrosomes are autocatalytic droplets of pericentriolar material organized by centrioles. PNAS. 2014 Jul 1;111(26):E2636-45. [PubMed]
      This paper proposes a model in which centrosomes are liquid droplets, formed when centrosomal material undergoes phase separation from the cytosol.
    • Goehring NW, Hyman AA. Organelle Growth Control through Limiting Pools of Cytoplasmic Components. Curr Biol. 2012 May 8;22(9):R330-9.  [PubMed]
      A review on the scaling of intracellular organelles, including the centrosome.
    • Decker M, et al. Limiting Amounts of Centrosome Material Set Centrosome Size in C. elegans Embryos. Curr Biol 21(15):1259-67 (2011).
      [PubMed]
      Centrosome size scales with cell size and inversely with cell number. Here the centrosome protein SPD-2 is identified as a limiting component for centrosome size, its’ concentration determining the size of the centrosomes inside the cell.
    • Jaensch S, Decker M, Hyman AA, & Myers EW. Automated tracking and analysis of centrosomes in early Caenorhabditis elegans embryos. Bioinformatics 26, i13–i20 (2010).
      [PubMed]
      Publication of a software capable of tracking centrosomes in C. elegans embryos in three dimensions throughout the development from the 1-cell to the 16-cell stage. Simultaneous automated data analysis yields valuable information about each tracked centrosome, like the trajectory or the size.
    • Greenan G, et al. Centrosome size sets mitotic spindle length in Caenorhabditis elegans embryos. Curr Biol 20, 353–358 (2010).
      [PubMed]
      The length of the mitotic spindle in C. elegans embryos directly correlates to the size of the centrosomes. Perturbation of centrosome size also changes the size of the spindle. This relation is connected to the centrosomes’ ability to recruit the microtubule associated protein TPXL-1.
    • Boxem M, et al. A protein domain-based interactome network for C. elegans early embryogenesis. Cell 134, 534–545 (2008).
      [PubMed]
      A protein-protein interaction screen for 800 proteins related to C. elegans early embryogenesis, yielding valuable information on the interactions of centrosome proteins as well as the architecture of the nuclear pore complex.
    • Pelletier L, O’toole E, Schwager A, Hyman AA, & Müller-Reichert T. Centriole assembly in Caenorhabditis elegans. Nature 444, 619–623 (2006).
      [PubMed]
      The pathway of centriole assembly starts with the SPD-2/ZYG-1 dependent recruitment of the SAS proteins to the mother centriole. Subsequently SAS-5/-6 ensure central tube formation, where as SAS-4 is required for the growth of the nine singlet microtubules surrounding the central tube.