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Publication : Coupling between clathrin-coated-pit invagination, cortactin recruitment, and membrane scission observed in live cells.

First Author  Merrifield CJ Year  2005
Journal  Cell Volume  121
Issue  4 Pages  593-606
PubMed ID  15907472 Mgi Jnum  J:98947
Mgi Id  MGI:3580762 Doi  10.1016/j.cell.2005.03.015
Citation  Merrifield CJ, et al. (2005) Coupling between clathrin-coated-pit invagination, cortactin recruitment, and membrane scission observed in live cells. Cell 121(4):593-606
abstractText  During clathrin-mediated endocytosis, membrane scission marks the isolation of a cargo-laden clathrin-coated pit (CCP) from the cell exterior. Here we used live-cell imaging of a pH-sensitive cargo to visualize the formation of clathrin-coated vesicles (CCVs) at single CCPs with a time resolution of seconds. We show that CCPs are highly dynamic and can produce multiple vesicles in succession. Using alternating evanescent field and epifluorescence illumination, we show that CCP invagination and scission are tightly coupled, with scission coinciding with maximal displacement of CCPs from the plasma membrane and with peak recruitment of cortactin-DsRed, a dynamin and F-actin binding protein. Finally, perturbing actin polymerization with latrunculin-B drastically reduces the efficiency of membrane scission and affects many aspects of CCP dynamics. We propose that CCP invagination, actin polymerization, and CCV formation are highly coordinated for efficient endocytosis.
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