GTP-binding proteins of the Sec4/Ypt1/rab subfamily of the Ras superfamily are thought to play an important role in controlling vesicular traffic along the secretory and endocytic pathways of all cells. The precise mechanism of action of these proteins remains to be elucidated. We propose here to study the role of rab proteins in the exo-endocytotic recycling of neuronal synaptic vesicles (SVs) and endocrine synaptic-like microvesicles (SLMVs). The large body of information already available on the biochemical composition of these organelles makes them optimal model systems to investigate the molecular mechanisms involved in membrane budding and membrane fusion. This project will be organized around four specific aims. First, the precise recycling pathway of SVs/SLMVs will be further investigated using a neuroendocrine cell line as a model system. Primary goal of this work is to test the hypothesis that SV/SLMV recycling involves transit through endosomes and sorting from endosomal membranes. Second, assays to study specific steps of the recycling of SLMVs will be developed. These will include assays to quantify vesicle exocytosis independent of neurotransmitter release. Third, these assays will be used to study the role of rab3a, rab4 and rab5 in specific steps of SLMV recycling. Fourth, the function in neurons and endocrine cells of the mammalian protein Mss4 which can suppress the secretory defect of the sec4-8 mutant of Saccaromyces cerevisiae will be investigated. The sec4-8 yeast strain harbors a point mutation in the GTP-binding protein Sec4 which is associated with secretory vesicles. Mss4 is a guanine nucleotide exchange protein homologous the yeast protein DSS4.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
3P01CA046128-14S1
Application #
6435825
Study Section
Project Start
2001-03-07
Project End
2001-12-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
14
Fiscal Year
2001
Total Cost
$197,196
Indirect Cost
Name
Yale University
Department
Type
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
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
Destaing, Olivier; Ferguson, Shawn M; Grichine, Alexei et al. (2013) Essential function of dynamin in the invasive properties and actin architecture of v-Src induced podosomes/invadosomes. PLoS One 8:e77956
Mao, Yuxin; Balkin, Daniel M; Zoncu, Roberto et al. (2009) A PH domain within OCRL bridges clathrin-mediated membrane trafficking to phosphoinositide metabolism. EMBO J 28:1831-42
Ferguson, Shawn M; Ferguson, Shawn; Raimondi, Andrea et al. (2009) Coordinated actions of actin and BAR proteins upstream of dynamin at endocytic clathrin-coated pits. Dev Cell 17:811-22
Chen, Hong; Ko, Genevieve; Zatti, Alessandra et al. (2009) Embryonic arrest at midgestation and disruption of Notch signaling produced by the absence of both epsin 1 and epsin 2 in mice. Proc Natl Acad Sci U S A 106:13838-43
Zoncu, Roberto; Perera, Rushika M; Balkin, Daniel M et al. (2009) A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes. Cell 136:1110-21
Gong, Liang-Wei; De Camilli, Pietro (2008) Regulation of postsynaptic AMPA responses by synaptojanin 1. Proc Natl Acad Sci U S A 105:17561-6
Frost, Adam; Perera, Rushika; Roux, Aurelien et al. (2008) Structural basis of membrane invagination by F-BAR domains. Cell 132:807-17
Hayashi, Mitsuko; Raimondi, Andrea; O'Toole, Eileen et al. (2008) Cell- and stimulus-dependent heterogeneity of synaptic vesicle endocytic recycling mechanisms revealed by studies of dynamin 1-null neurons. Proc Natl Acad Sci U S A 105:2175-80
Destaing, Olivier; Sanjay, Archana; Itzstein, Cecile et al. (2008) The tyrosine kinase activity of c-Src regulates actin dynamics and organization of podosomes in osteoclasts. Mol Biol Cell 19:394-404

Showing the most recent 10 out of 167 publications