This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Centrioles are organelles that play essential roles in the formation of cilia and flagella, and they participate in cytokinesis, cell cycle control and centrosome organization. The cell's single centrosome, which is inherited as a spindle pole, duplicates only once per cell cycle, a process that is regulated through the control of centriole duplication. Centriole duplication is strictly controlled to ensure the formation of a bipolar spindle and the maintenance of ploidy. It has been known for many years that daughter centrioles assemble at right angles to the older, or """"""""mother"""""""" centriole. The significance of this and the pathway for daughter centriole assemble remain mysterious. We are using the correlative light and EM tomography approach to study centrosome assembly in staged C. elegans embryos. Our initial results showed that daughter centriole assembly in this organism begins with the formation of a central tube, just as the pronuclei appear (Pelletier, L., O'Toole, E.T., Schwager, A., Hyman, A.A., and M?ller-Reichert, T. (2006) Centriole assembly in Caenorhabditis elegans. Nature 444, 619-23). The central tube elongates during pronuclear migration, and the assembly of singlet microtubules on the tube occurs during pronuclear rotation. Using EM-tomography in combination with RNA interference and restoration microscopy we have found evidence suggesting that tube formation and elongation requires the SAS-5/SAS-6 proteins and an upstream signal mediated by SPD-2 and ZYG-1. We further show that although the assembly of the central tube still occurs, singlet microtubules fail to assemble in the absence of SAS-4, suggesting that it is required specifically at that step of assembly. The centriole assembly pathway we describe here in C. elegans may provide a framework for further studies on centriole assembly in other centriole bearing organisms.

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-CB-J (40))
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Colorado at Boulder
Schools of Arts and Sciences
United States
Zip Code
Giddings Jr, Thomas H; Morphew, Mary K; McIntosh, J Richard (2017) Preparing Fission Yeast for Electron Microscopy. Cold Spring Harb Protoc 2017:pdb.prot091314
Zhao, Xiaowei; Schwartz, Cindi L; Pierson, Jason et al. (2017) Three-Dimensional Structure of the Ultraoligotrophic Marine Bacterium ""Candidatus Pelagibacter ubique"". Appl Environ Microbiol 83:
Brown, Joanna R; Schwartz, Cindi L; Heumann, John M et al. (2016) A detailed look at the cytoskeletal architecture of the Giardia lamblia ventral disc. J Struct Biol 194:38-48
Saheki, Yasunori; Bian, Xin; Schauder, Curtis M et al. (2016) Control of plasma membrane lipid homeostasis by the extended synaptotagmins. Nat Cell Biol 18:504-15
Höög, Johanna L; Lacomble, Sylvain; Bouchet-Marquis, Cedric et al. (2016) 3D Architecture of the Trypanosoma brucei Flagella Connector, a Mobile Transmembrane Junction. PLoS Negl Trop Dis 10:e0004312
Park, J Genevieve; Palmer, Amy E (2015) Properties and use of genetically encoded FRET sensors for cytosolic and organellar Ca2+ measurements. Cold Spring Harb Protoc 2015:pdb.top066043
McCoy, Kelsey M; Tubman, Emily S; Claas, Allison et al. (2015) Physical limits on kinesin-5-mediated chromosome congression in the smallest mitotic spindles. Mol Biol Cell 26:3999-4014
Höög, Johanna L; Lötvall, Jan (2015) Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy. J Extracell Vesicles 4:28680
Marc, Robert E; Anderson, James R; Jones, Bryan W et al. (2014) The AII amacrine cell connectome: a dense network hub. Front Neural Circuits 8:104
Weber, Britta; Tranfield, Erin M; Höög, Johanna L et al. (2014) Automated stitching of microtubule centerlines across serial electron tomograms. PLoS One 9:e113222

Showing the most recent 10 out of 84 publications