Cells require ordered movement of proteins and lipids from one membrane-bound compartment to another, while maintaining the organization, function and heterogeneity of the donor and acceptor membranes;vesicular membrane traffic carries out these functions. Clathrin coated vesicles are the most prominent form of traffic from the plasma membrane to endosomes (endocytosis), a pathway by which various ligands and their receptors enter cells. Clathrin coated vesicles are also important for traffic between endosomes and the trans-Golgi network (TGN). We propose to continue our studies of molecular mechanisms in clathrin-mediated membrane traffic, using direct observation by live-cell imaging with single-molecular sensitivity and high spatial and temporal resolution, to answer the following questions. (1) What are the molecular events required to initiate formation of a clathrin coated pit? (2) What are the molecular components of coats trapped at various stages of growth and closure: early and late abortive coated pits, and pits stalled at a relatively late stage, by imposing tension of the membrane and by inhibiting actin dynamics? (3) What is the mechanism of clathrin uncoating mediated by Hsc70 and its co-chaperone auxilin and how is the uncoating coordinated with assembly so that the two processes do not compete?

Public Health Relevance

Vesicular membrane traffic is the principal mechanism for moving proteins and lipids among membrane-bound compartments in a cell. Clathrin coated vesicles are the most prominent form of traffic from the plasma membrane to endosomes (endocytosis), a pathway by which ligands such as hormones, transferrin, immunoglobulins, LDL, viruses and their receptors enter cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM075252-08
Application #
8392265
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Ainsztein, Alexandra M
Project Start
2005-09-10
Project End
2014-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
8
Fiscal Year
2013
Total Cost
$376,350
Indirect Cost
$160,057
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Coulter, Michael E; Dorobantu, Cristina M; Lodewijk, Gerrald A et al. (2018) The ESCRT-III Protein CHMP1A Mediates Secretion of Sonic Hedgehog on a Distinctive Subtype of Extracellular Vesicles. Cell Rep 24:973-986.e8
Swinburne, Ian A; Mosaliganti, Kishore R; Upadhyayula, Srigokul et al. (2018) Lamellar projections in the endolymphatic sac act as a relief valve to regulate inner ear pressure. Elife 7:
Liu, Tsung-Li; Upadhyayula, Srigokul; Milkie, Daniel E et al. (2018) Observing the cell in its native state: Imaging subcellular dynamics in multicellular organisms. Science 360:
Schuler, Max-Hinderk; Lewandowska, Agnieszka; Caprio, Giuseppe Di et al. (2017) Miro1-mediated mitochondrial positioning shapes intracellular energy gradients required for cell migration. Mol Biol Cell 28:2159-2169
Adell, Manuel Alonso Y; Migliano, Simona M; Upadhyayula, Srigokul et al. (2017) Recruitment dynamics of ESCRT-III and Vps4 to endosomes and implications for reverse membrane budding. Elife 6:
He, Kangmin; Marsland III, Robert; Upadhyayula, Srigokul et al. (2017) Dynamics of phosphoinositide conversion in clathrin-mediated endocytic traffic. Nature 552:410-414
Chou, Yi-Ying; Cuevas, Christian; Carocci, Margot et al. (2016) Identification and Characterization of a Novel Broad-Spectrum Virus Entry Inhibitor. J Virol 90:4494-4510
Hu, Yu; Kim, Ji Hyung; He, Kangmin et al. (2016) Scramblase TMEM16F terminates T cell receptor signaling to restrict T cell exhaustion. J Exp Med 213:2759-2772
Aguet, François; Upadhyayula, Srigokul; Gaudin, Raphaël et al. (2016) Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy. Mol Biol Cell 27:3418-3435
Reeves, Patrick M; Kang, Yuan-Lin; Kirchhausen, Tom (2016) Endocytosis of Ligand-Activated Sphingosine 1-Phosphate Receptor 1 Mediated by the Clathrin-Pathway. Traffic 17:40-52

Showing the most recent 10 out of 65 publications