The goal of this proposal is to understand the mechanism by which eukaryotic cells target new membrane growth and secretion to specific sites on the cell surface and how this is coordinated with changes in cell polarity during the cell cycle. In this proposal we will examine the specific role of Lgl/Sro7 and Rho/Cdc42 protein families in this process. The Lgl/Sro7 family of proteins, first.identified as a tumor suppressor in Drosophila, is found in all eukaryotic cells and is likely to have a highly conserved function in regulation of polarity and exocytosis. We have characterized homologs of this family in both yeast and mammalian epithelial cells where they are found in physical association with a specific set of SNARE proteins. We will test the hypothesis that Lgl family members have a conserved structure and function in polarized exocytosis and may act as effectors of Rab GTPases in mediating SNARE-dependent vesicle fusion during polarized growth. Genetic analyses performed in our lab have led to the identification of a direct role for Rho/Cdc42 GTPases in exocytosis. In particular we have found two Rho GTPases, Rho3 and Cdc42, that function in a pathway which involves spatial regulation of exocytosis through direct activation of a multisubunit complex known as the Exocyst. In this proposal we will take a comprehensive approach to identify the molecular mechanism by which Sro7, Rho3, and Cdc42 function in the regulation of polarized exocytosis. Ultimately understanding the molecular details of these processes may allow the development of new approaches and novel therapeutics to combating cancer, or type II diabetes, as well as other diseases in which regulation of cell surface trafficking is central to the etiology of the disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM054712-14
Application #
7827961
Study Section
Special Emphasis Panel (ZRG1-CB-G (02))
Program Officer
Ainsztein, Alexandra M
Project Start
1998-05-01
Project End
2012-03-31
Budget Start
2010-05-01
Budget End
2012-03-31
Support Year
14
Fiscal Year
2010
Total Cost
$360,477
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Physiology
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Rossi, Guendalina; Watson, Kelly; Demonch, Mallory et al. (2015) In vitro reconstitution of Rab GTPase-dependent vesicle clustering by the yeast lethal giant larvae/tomosyn homolog, Sro7. J Biol Chem 290:612-24
Watson, Kelly; Rossi, Guendalina; Temple, Brenda et al. (2015) Structural basis for recognition of the Sec4 Rab GTPase by its effector, the Lgl/tomosyn homologue, Sro7. Mol Biol Cell 26:3289-300
Watson, Leah J; Rossi, Guendalina; Brennwald, Patrick (2014) Quantitative analysis of membrane trafficking in regulation of Cdc42 polarity. Traffic 15:1330-43
Brennwald, Patrick (2013) Membrane traffic: the exocyst meets the cell cycle. Curr Biol 23:R838-40
Forsmark, Annabelle; Rossi, Guendalina; Wadskog, Ingrid et al. (2011) Quantitative proteomics of yeast post-Golgi vesicles reveals a discriminating role for Sro7p in protein secretion. Traffic 12:740-53
Rossi, Guendalina; Brennwald, Patrick (2011) Yeast homologues of lethal giant larvae and type V myosin cooperate in the regulation of Rab-dependent vesicle clustering and polarized exocytosis. Mol Biol Cell 22:842-57
Boulter, Etienne; Garcia-Mata, Rafael; Guilluy, Christophe et al. (2010) Regulation of Rho GTPase crosstalk, degradation and activity by RhoGDI1. Nat Cell Biol 12:477-83
Wu, Hao; Turner, Courtney; Gardner, Jimmy et al. (2010) The Exo70 subunit of the exocyst is an effector for both Cdc42 and Rho3 function in polarized exocytosis. Mol Biol Cell 21:430-42
Wu, Hao; Brennwald, Patrick (2010) The function of two Rho family GTPases is determined by distinct patterns of cell surface localization. Mol Cell Biol 30:5207-17
Ivanov, Andrei I; Young, Cheryl; Den Beste, Kyle et al. (2010) Tumor suppressor scribble regulates assembly of tight junctions in the intestinal epithelium. Am J Pathol 176:134-45

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