Molecular analysis of the Yeast Actin Cytoskeleton We will use biochemical and genetic approaches to study actin assembly in yeast. These studies will help to elucidate the molecular mechanisms responsible for cellular morphogenesis. The sophisticated classical and molecular genetics of the yeast Saccharomyces cerevisiae, together with there simplicity of the organism compared to higher cells, make it particularly tractable for these studies. These specific questions will be addressed: 1. How does actin assemble? The assembly properties of wild type and mutant actins will be determined in vitro to provide functional tests of the molecular model of the actin filament. In addition, phenotypic analysis of a full spectrum of actin mutants will provide a better definition of there in vivo roles of actin. 2. What are the roles of actin-binding proteins? The two yeast actin-binding proteins encoded by the SAC6 and ABP1 genes will be studied biochemically and genetically to gain an understanding of these proteins at a level not yet achieved for actin-binding proteins in previous studies using genetically intractable organisms. The function an ABP1 protein domain homologous to tyrosine kinase proto-oncogene regulatory domains will be explored in detail. 3. How is the organization of the actin cytoskeleton controlled spatially and temporally? Biochemical and genetic approaches will be used to identify structural proteins that bind to and organize the actin cytoskeleton. Classical genetic approaches will be used to identify the proteins that transduce regulatory signals to the actin cytoskeleton, and second messenger molecules that can affect actin-binding protein activities will be identified using biochemical assays. In total, these studies will address central questions about the function of the actin cytoskeleton in vivo and about how the cytoskeleton is organized; the disposition of the cytoskeleton within the cell is central to its function. What is learned from these studies is likely to apply to higher cells where defective actin cytoskeleton function is implicated in pathological conditions such as muscular dystrophy and neoplastic transformation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042759-04
Application #
3301601
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1989-07-01
Project End
1994-06-30
Budget Start
1992-07-01
Budget End
1993-06-30
Support Year
4
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Type
Schools of Arts and Sciences
DUNS #
094878337
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Lu, Rebecca; Drubin, David G; Sun, Yidi (2016) Clathrin-mediated endocytosis in budding yeast at a glance. J Cell Sci 129:1531-6
Miao, Yansong; Han, Xuemei; Zheng, Liangzhen et al. (2016) Fimbrin phosphorylation by metaphase Cdk1 regulates actin cable dynamics in budding yeast. Nat Commun 7:11265
Sun, Yidi; Leong, Nicole T; Wong, Tiffany et al. (2015) A Pan1/End3/Sla1 complex links Arp2/3-mediated actin assembly to sites of clathrin-mediated endocytosis. Mol Biol Cell 26:3841-56
Lewellyn, Eric B; Pedersen, Ross T A; Hong, Jessica et al. (2015) An Engineered Minimal WASP-Myosin Fusion Protein Reveals Essential Functions for Endocytosis. Dev Cell 35:281-94
Cortesio, Christa L; Lewellyn, Eric B; Drubin, David G (2015) Control of lipid organization and actin assembly during clathrin-mediated endocytosis by the cytoplasmic tail of the rhomboid protein Rbd2. Mol Biol Cell 26:1509-22
Michelot, Alphée; Drubin, David G (2014) Dissecting principles governing actin assembly using yeast extracts. Methods Enzymol 540:381-97
Weinberg, Jasper S; Drubin, David G (2014) Regulation of clathrin-mediated endocytosis by dynamic ubiquitination and deubiquitination. Curr Biol 24:951-9
Michelot, Alphée; Grassart, Alexandre; Okreglak, Voytek et al. (2013) Actin filament elongation in Arp2/3-derived networks is controlled by three distinct mechanisms. Dev Cell 24:182-95
Miao, Yansong; Wong, Catherine C L; Mennella, Vito et al. (2013) Cell-cycle regulation of formin-mediated actin cable assembly. Proc Natl Acad Sci U S A 110:E4446-55
Weinberg, Jasper; Drubin, David G (2012) Clathrin-mediated endocytosis in budding yeast. Trends Cell Biol 22:1-13

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