Abstract
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 α factor receptor, Ste2p. Cortical clathrin localizes with epsins Ent1/2p 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.
| Original language | English |
|---|---|
| Pages (from-to) | 87-98 |
| Number of pages | 12 |
| Journal | Developmental Cell |
| Volume | 9 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jul 1 2005 |
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- Developmental Biology
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In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast. / Newpher, Thomas M.; Smith, Robin P.; Lemmon, Vance; Lemmon, Sandra.
In: Developmental Cell, Vol. 9, No. 1, 01.07.2005, p. 87-98.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast
AU - Newpher, Thomas M.
AU - Smith, Robin P.
AU - Lemmon, Vance
AU - Lemmon, Sandra
PY - 2005/7/1
Y1 - 2005/7/1
N2 - 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 α factor receptor, Ste2p. Cortical clathrin localizes with epsins Ent1/2p 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.
AB - 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 α factor receptor, Ste2p. Cortical clathrin localizes with epsins Ent1/2p 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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U2 - 10.1016/j.devcel.2005.04.014
DO - 10.1016/j.devcel.2005.04.014
M3 - Article
VL - 9
SP - 87
EP - 98
JO - Developmental Cell
JF - Developmental Cell
SN - 1534-5807
IS - 1
ER -