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 K.
N1 - Funding Information:
We thank David Drubin, Beverly Wendland, Erin O’Shea, Kenneth Blumer, and the Yeast Resource Center for providing many strains and plasmids. We thank Kenneth Henry, Ji Suk Chang, and John Collette for many helpful discussions. T.M.N. was supported by NIH training grant (T32 GM08056-20). This work was funded by NIH grants R01 GM55796 (S.K.L.) and R01 EY05285 (V.L.).
PY - 2005/7
Y1 - 2005/7
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.
UR - http://www.scopus.com/inward/record.url?scp=21344469702&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=21344469702&partnerID=8YFLogxK
U2 - 10.1016/j.devcel.2005.04.014
DO - 10.1016/j.devcel.2005.04.014
M3 - Article
C2 - 15992543
AN - SCOPUS:21344469702
VL - 9
SP - 87
EP - 98
JO - Developmental Cell
JF - Developmental Cell
SN - 1534-5807
IS - 1
ER -