Internalization of integral membrane proteins during receptor- mediated endocytosis (RME) is accompanied by the polymerization of clathrin on the cytoplasmic side of invaginated membrane. This results in the formation of coated pits and coated vesicles, composed of internalized membrane and receptors, covered with a polyhedral lattice of clathrin. It has been proposed that self-assembly of clathrin into this polyhedral structure is fundamental to the process of selective receptor internalization and membrane translocation which occurs during RME. To understand the mechanism and ultimately the control of this selective membrane movement, we propose to investigate the molecular basis of clathrin polymerization both in vitro and in situ. In preliminary studies, we have identifed molecular sites involved in clathrin-clathrin interactions during assembly by using anti-clathrin monoclonal antibodies (MAbs) to inhibit clathrin assembly. Based on the aborted clathrin structures produced, we have proposed a model for clathrin assembly which involves sequential molecular interactions. The first specific aim of the studies outlined in this proposal is to focus on molecular localization of clathrin interaction sites involved in assembly and to test the proposed model. The second specific aim is to address the role of accessory proteins in clathrin polymerization to assess additional fators which influence clathrin assembly in situ and affect coated vesicle movement. The third specific aim is to investigae in situ, clathrin assembly by following biosynthesis and assembly of the clathrin subunits. As a whole, these studies should lead to a clearer understanding of the process of clathrin assembly at the molecular and cellular level and establish directions for investigating additional processes contributing to receptor-mediated endocytosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038093-02
Application #
3294122
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1988-02-01
Project End
1991-01-31
Budget Start
1989-02-01
Budget End
1990-01-31
Support Year
2
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Pharmacy
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
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Goyos, Ana; Guethlein, Lisbeth A; Horowitz, Amir et al. (2015) A Distinctive Cytoplasmic Tail Contributes to Low Surface Expression and Intracellular Retention of the Patr-AL MHC Class I Molecule. J Immunol 195:3725-36
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Majeed, Sophia R; Vasudevan, Lavanya; Chen, Chih-Ying et al. (2014) Clathrin light chains are required for the gyrating-clathrin recycling pathway and thereby promote cell migration. Nat Commun 5:3891
Young, Anna; Stoilova-McPhie, Svetla; Rothnie, Alice et al. (2013) Hsc70-induced changes in clathrin-auxilin cage structure suggest a role for clathrin light chains in cage disassembly. Traffic 14:987-96
Hoshino, Sachiko; Sakamoto, Kazuho; Vassilopoulos, Stéphane et al. (2013) The CHC22 clathrin-GLUT4 transport pathway contributes to skeletal muscle regeneration. PLoS One 8:e77787
Holmes, Brandon B; DeVos, Sarah L; Kfoury, Najla et al. (2013) Heparan sulfate proteoglycans mediate internalization and propagation of specific proteopathic seeds. Proc Natl Acad Sci U S A 110:E3138-47
Asensio, Cedric S; Sirkis, Daniel W; Maas Jr, James W et al. (2013) Self-assembly of VPS41 promotes sorting required for biogenesis of the regulated secretory pathway. Dev Cell 27:425-37

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