Journal
DEVELOPMENTAL CELL
Volume 9, Issue 1, Pages 87-98Publisher
CELL PRESS
DOI: 10.1016/j.devcel.2005.04.014
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Funding
- NEI NIH HHS [R01 EY05285, R01 EY005285] Funding Source: Medline
- NIGMS NIH HHS [R01 GM55796, T32 GM08056-20] Funding Source: Medline
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Clathrin-mediated transport is a major pathway for endocytosis. However, in yeast, where cortical actin patches are essential for endocytosis, plasma membrane-associated clathrin has never been observed. Using live cell imaging, we demonstrate cortical clathrin in association with the actin-based endocytic machinery in yeast. Fluorescently tagged clathrin is found in highly mobile internal trans-Golgi/endosomal structures and in smaller cortical patches. Total internal reflection fluorescence microscopy showed that cortical patches are likely endocytic sites, as clathrin is recruited prior to a burst of intensity of the actin patch/endocytic marker, Abp1. Clathrin also accumulates at the cortex with internalizing a factor receptor, Ste2p. Cortical clathrin localizes with epsins Entl/2p and AP180s, and its recruitment to the surface is dependent upon these adaptors. In contrast, Sla2p, End3p, Pan1p, and a dynamic actin cytoskeleton are not required for clathrin assembly or exchange but are required for the mobility, maturation, and/or turnover of clathrin-containing endocytic structures.
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