The fine structure of Schizosaccharomyces pombe has been preserved with unprecedented quality, using a high pressure freezer and freeze substitution fixation. The resulting EM specimens have allowed us both to analyze the 3-D fine structure of the mitotic spindles in S. pombe (Ding et al. 1993) and to investigate the structure of specific intracellular organelles, such as the spindle pole bodies (SPBs). We have now characterized the process of SPB duplication in wild-type cells. During interphase, the SPB resides in the cytoplasm, immediately associated with the nuclear envelope. It duplicates late in G2, but the two SPBs remain attached as they settle into an invagination that forms in the nuclear envelope. This membrane specialization then opens to form a special fenestra in the nuclear envelope. The duplicated SPB attaches to the edges of the fenestra, then MTs grow from the inner surface of the SPBs as they separate and move to become the poles of the forming spindle. During anaphase, the SPBs are extruded from the fenestrae at each end of the spindle and re-enter the cytoplasm, where they reside until they duplicate and repeat the cycle (Ding et al., Mol Biol. Cell). We have also fixed various strains of yeast that are mutant in genes required for normal mitosis in order to use 3-D fine structural analysis to characterize mutant phenotype and to help us understand the function of the corresponding wild type gene products in the normal cell division process. Seven cells carrying the the mitotic mutation, cut11-2 have been reconstructed in 3-D, allowing us to learn that these cells have aberrant spindle pole function. The Cut11 protein product appears to be required for normal attachment of the SPB to the nuclear envelope during mitosis (West et al., in preparation). ?
| Giddings Jr, Thomas H; Morphew, Mary K; McIntosh, J Richard (2017) Preparing Fission Yeast for Electron Microscopy. Cold Spring Harb Protoc 2017: |
| 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
