This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In budding yeast, the spindle pole body (SPB) is the sole microtubule organizing center of the cell. During meiosis, however, the cytoplasmic face of the SPB is converted from a site of microtubule nucleation to a site of membrane formation. This transformation is accomplished by the recruitment of at least four meiosis-specific proteins that displace the gamma-tubulin complex and form a novel structure, termed the meiotic outer plaque (MOP), which serves as a site of de novo membrane formation. The MOP is a highly ordered structure and several proteins that localize to the MOP have been identified, but how they are arranged and how they promote membrane formation is not known. We have immuno labeled strains of S. cerevisiae that have different spindle pole body proteins fused with GFP. Both Ady4p and Spo74p localize to the MOP and have been shown to promote prospore membrane growth. The goal of this project is to develop a high-resolution structural model of this membrane organizing center. The small size of the complex (300-400nm in diameter) make Immuno-EM and EM tomography essential for this analysis. By combining these techniques with mutants lacking individual protein components of the MOP, as well as results from fluorescence-based analysis, a robust model of the organization of the MOP will be generated. Such a model will be an important step in the long-term goal of understanding how the MOP promotes the formation of intracellular membranes. We have begun to characterize the first stages of pro-spore membrane (PSM) formation in wild type cells as well as the lack of PSM formation in Spo14 deletion mutants.
| 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: |
| 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 |
| 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 |
| 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
