The goal of this research program is to understand clathrin biochemistry in order to elucidate how clathrin- coated vesicle (CCV) formation and function is regulated in cells. CCVs implement the fundamental cellular membrane traffic pathways of endocytosis and lysosome biogenesis. CCVs also participate in specialized protein sorting pathways in specific tissues, influencing physiological pathways that play critical roles in human health. These include nutrient and cholesterol uptake, glucose transporter and insulin targeting, immune receptor traffic during antigen presentation and synaptic vesicle protein recapture. The characteristic lattice structure of the CCV coat that is responsible for protein sorting is formed by interactions between clathrin heavy chains (CHCs). Clathrin light chain (CLC) subunits mediate CCV binding to Hip proteins (Hip1 and Hip1R), which can bind actin and cortactin and thereby regulate actin dynamics in the vicinity of the clathrin lattice. Work from the past funding period has defined through structural and biochemical studies how CLCs also contribute to regulation of clathrin lattice formation. Now that these biochemical properties of CLCs have been established, it remains to be determined how they influence clathrin function in cells and tissues and to discover the unique functions of the two different forms of CLC (LCa and LCb). Understanding these aspects of CLC function will be the main focus for the next funding period, as this is a remaining frontier for establishing molecular mechanisms that control specificity of clathrin pathways and diversify their cellular function. Experiments to improve structural characterization of CCV proteins and analysis of how signaling regulates CHC function will be continued from the last funding period. With these goals, the three specific aims for the next funding period are as follows.
Aim 1 is to determine the molecular mechanisms for intrinsic control of clathrin function through further structural analysis of clathrin and Hip proteins and through biochemical and cellular analysis of clathrin heavy chain phosphorylation.
Aim 2 is to elucidate the role of clathrin light chains and their interaction with Hip proteins in cellular functions, including migration.
Aim 3 is to establish the relative roles of the two vertebrate clathrin light chains LCa and LCb at the tissue level through targeted gene deletion from mice. The experimental approaches to these studies include protein crystallography and structure-based mutagenesis, RNA interference, membrane traffic and migration assays, and gene targeting strategies to produce animals that can be conditionally depleted of LCa or LCb in specific tissues, followed by analysis of immune function and neurological function. These studies continue to pursue our long-term strategy for understanding complex cellular pathways of clathrin function that are relevant to human health and disease through applying knowledge of clathrin biochemistry to focused studies of cells, tissues and organisms.

Public Health Relevance

The clathrin protein is involved in key cellular transport pathways that contribute to maintenance of human health including nutrition, lipid metabolism, hormone regulation, the immune response and cell growth control. Understanding clathrin function therefore has relevance for establishing molecular mechanisms of a number of human disease states such as heart disease, diabetes, cancer, neuro-muscular defects, and bacterial and viral infection.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
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Ainsztein, Alexandra M
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University of California San Francisco
Schools of Pharmacy
San Francisco
United States
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Vassilopoulos, Stéphane; Gentil, Christel; Lainé, Jeanne et al. (2014) Actin scaffolding by clathrin heavy chain is required for skeletal muscle sarcomere organization. J Cell Biol 205:377-93
Sullivan, Chelsea S; Scheib, Jami L; Ma, Zhong et al. (2014) The adaptor protein GULP promotes Jedi-1-mediated phagocytosis through a clathrin-dependent mechanism. Mol Biol Cell 25:1925-36
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
Brodsky, Frances M; Sosa, R Thomas; Ybe, Joel A et al. (2014) Unconventional functions for clathrin, ESCRTs, and other endocytic regulators in the cytoskeleton, cell cycle, nucleus, and beyond: links to human disease. Cold Spring Harb Perspect Biol 6:a017004
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
Stachowiak, Jeanne C; Brodsky, Frances M; Miller, Elizabeth A (2013) A cost-benefit analysis of the physical mechanisms of membrane curvature. Nat Cell Biol 15:1019-27
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
Stoddart, A; Tennant, T R; Fernald, A A et al. (2012) The clathrin-binding domain of CALM-AF10 alters the phenotype of myeloid neoplasms in mice. Oncogene 31:494-506
Shieh, Jennifer C; Schaar, Bruce T; Srinivasan, Karpagam et al. (2011) Endocytosis regulates cell soma translocation and the distribution of adhesion proteins in migrating neurons. PLoS One 6:e17802
Bonazzi, Matteo; Vasudevan, Lavanya; Mallet, Adeline et al. (2011) Clathrin phosphorylation is required for actin recruitment at sites of bacterial adhesion and internalization. J Cell Biol 195:525-36

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