In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast

Thomas M. Newpher, Robin P. Smith, Vance Lemmon, Sandra Lemmon

Research output: Contribution to journalArticle

137 Citations (Scopus)

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 languageEnglish
Pages (from-to)87-98
Number of pages12
JournalDevelopmental Cell
Volume9
Issue number1
DOIs
StatePublished - Jul 1 2005

Fingerprint

Vesicular Transport Adaptor Proteins
Clathrin
Yeast
Machinery
Actins
Yeasts
Endocytosis
Fluorescence microscopy
Cell membranes
Actin Cytoskeleton
Fluorescence Microscopy
Cell Membrane

ASJC Scopus subject areas

  • Developmental Biology

Cite this

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 journalArticle

@article{e2345b4e2e4d4109955b01b8213e6af3,
title = "In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast",
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.",
author = "Newpher, {Thomas M.} and Smith, {Robin P.} and Vance Lemmon and Sandra Lemmon",
year = "2005",
month = "7",
day = "1",
doi = "10.1016/j.devcel.2005.04.014",
language = "English",
volume = "9",
pages = "87--98",
journal = "Developmental Cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "1",

}

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.

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

VL - 9

SP - 87

EP - 98

JO - Developmental Cell

JF - Developmental Cell

SN - 1534-5807

IS - 1

ER -