Recent improvements in electron microscopes in equipment for digital image processing imaging processing, and in methods for biological specimen preparation and analysis have set the stage for a revolution in cellular electron microscopy. This Program Project assembles cell biologists at the University of Colorado with diverse scientific and technological experience so they can make a contribution to this will be significant. Our program project includes a major section on Core Technological Research and Development in which we will implement highly efficient EM tomography of significant volumes from fast frozen, freeze-substituted cells whose preservation for structural study represents the state-of-the art. Core projects include novel approaches to the labeling of specific cellular macromolecules, innovations in structural modeling technology, and new approaches to the labeling of specific cellular macromolecules, innovations in structural modeling technology, and new approaches to thinking about the structural variability of organelles. Three scientific projects will capitalize on this technology and contribute to its development: an analysis of Golgi structure and function in mammalian cells; descriptions of Golgi variability and complexity in yeast, algae, and plants; a structural dissection of the centrosome from budding yeast; and an experimental/descriptive analysis of mammalian kinetochores. Each of these projects will require the instrumentation and technology developed in the Core, but at the same time each scientific project will provide a viable test bed for the efficacy of the new technologies developed. The members of each project are accustomed to collaborative effort, and they see this integrated approach to improving the study of cellular structure as a major project for their coming 5 years of research.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM061306-02
Application #
6387161
Study Section
Special Emphasis Panel (ZRG1-SSS-I (02))
Program Officer
Deatherage, James F
Project Start
2000-05-01
Project End
2005-04-30
Budget Start
2001-05-01
Budget End
2002-04-30
Support Year
2
Fiscal Year
2001
Total Cost
$1,026,514
Indirect Cost
Name
University of Colorado at Boulder
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
City
Boulder
State
CO
Country
United States
Zip Code
80309
Gergely, Zachary R; Martinez, Dana E; Donohoe, Bryon S et al. (2018) 3D electron tomographic and biochemical analysis of ER, Golgi and trans Golgi network membrane systems in stimulated Venus flytrap (Dionaea muscipula) glandular cells. J Biol Res (Thessalon) 25:15
Donohoe, Bryon S; Kang, Byung-Ho; Gerl, Mathias J et al. (2013) Cis-Golgi cisternal assembly and biosynthetic activation occur sequentially in plants and algae. Traffic 14:551-67
Zheng, Huiqiong; Staehelin, L Andrew (2011) Protein storage vacuoles are transformed into lytic vacuoles in root meristematic cells of germinating seedlings by multiple, cell type-specific mechanisms. Plant Physiol 155:2023-35
Kang, Byung-Ho; Nielsen, Erik; Preuss, Mary Lai et al. (2011) Electron tomography of RabA4b- and PI-4Kýý1-labeled trans Golgi network compartments in Arabidopsis. Traffic 12:313-29
Segui-Simarro, Jose M; Staehelin, L Andrew (2009) Mitochondrial reticulation in shoot apical meristem cells of Arabidopsis provides a mechanism for homogenization of mtDNA prior to gamete formation. Plant Signal Behav 4:168-71
Karahara, Ichirou; Suda, Jinsuke; Tahara, Hiroshi et al. (2009) The preprophase band is a localized center of clathrin-mediated endocytosis in late prophase cells of the onion cotyledon epidermis. Plant J 57:819-31
Limbach, Christoph; Staehelin, L Andrew; Sievers, Andreas et al. (2008) Electron tomographic characterization of a vacuolar reticulum and of six vesicle types that occupy different cytoplasmic domains in the apex of tip-growing Chara rhizoids. Planta 227:1101-14
Staehelin, L Andrew; Kang, Byung-Ho (2008) Nanoscale architecture of endoplasmic reticulum export sites and of Golgi membranes as determined by electron tomography. Plant Physiol 147:1454-68
Kang, Byung-Ho; Staehelin, L Andrew (2008) ER-to-Golgi transport by COPII vesicles in Arabidopsis involves a ribosome-excluding scaffold that is transferred with the vesicles to the Golgi matrix. Protoplasma 234:51-64
Segui-Simarro, Jose M; Coronado, Maria Jose; Staehelin, L Andrew (2008) The mitochondrial cycle of Arabidopsis shoot apical meristem and leaf primordium meristematic cells is defined by a perinuclear tentaculate/cage-like mitochondrion. Plant Physiol 148:1380-93

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