ADP-ribosylation factor 1 dependent clathrin-coat assembly on synthetic liposomes

Yunxiang Zhu, Matthew M Drake, Stuart Kornfeld

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The assembly of clathrin-coated vesicles on Golgi membranes is initiated by the GTP-binding protein ADP ribosylation factor (ARF), which generates high-affinity membrane-binding sites for the heterotetrameric AP-1 adaptor complex. Once bound, the AP-1 recruits clathrin triskelia, which polymerize to form the coat. We have found that ARF·GTP also recruits AP-1 and clathrin onto protein-free liposomes. The efficiency of this process is modulated by the composition of the liposomes, with phosphatidylserine being the most stimulatory phospholipid. There is also a requirement for cytosolic factor(s) other than ARF. Thin-section electron microscopy shows the presence of clathrin-coated buds and vesicles that resemble those formed in vivo. These results indicate that AP-1-containing clathrin-coated vesicles can form in the absence of integral membrane proteins. Thus, ARF·GTP, appropriate lipids, and cytosolic factor(s) are the minimal components necessary for AP- 1 clathrin-coat assembly.

Original languageEnglish (US)
Pages (from-to)5013-5018
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number9
DOIs
StatePublished - Apr 27 1999
Externally publishedYes

Fingerprint

ADP-Ribosylation Factor 1
Clathrin
Transcription Factor AP-1
Liposomes
Clathrin-Coated Vesicles
ADP-Ribosylation Factors
Membranes
Phosphatidylserines
GTP-Binding Proteins
Phospholipids
Electron Microscopy
Membrane Proteins
Binding Sites
Lipids

Keywords

  • AP-1 adaptor complex

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

ADP-ribosylation factor 1 dependent clathrin-coat assembly on synthetic liposomes. / Zhu, Yunxiang; Drake, Matthew M; Kornfeld, Stuart.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 9, 27.04.1999, p. 5013-5018.

Research output: Contribution to journalArticle

@article{8ae6849cec8348e9b367ac608daa3fd8,
title = "ADP-ribosylation factor 1 dependent clathrin-coat assembly on synthetic liposomes",
abstract = "The assembly of clathrin-coated vesicles on Golgi membranes is initiated by the GTP-binding protein ADP ribosylation factor (ARF), which generates high-affinity membrane-binding sites for the heterotetrameric AP-1 adaptor complex. Once bound, the AP-1 recruits clathrin triskelia, which polymerize to form the coat. We have found that ARF·GTP also recruits AP-1 and clathrin onto protein-free liposomes. The efficiency of this process is modulated by the composition of the liposomes, with phosphatidylserine being the most stimulatory phospholipid. There is also a requirement for cytosolic factor(s) other than ARF. Thin-section electron microscopy shows the presence of clathrin-coated buds and vesicles that resemble those formed in vivo. These results indicate that AP-1-containing clathrin-coated vesicles can form in the absence of integral membrane proteins. Thus, ARF·GTP, appropriate lipids, and cytosolic factor(s) are the minimal components necessary for AP- 1 clathrin-coat assembly.",
keywords = "AP-1 adaptor complex",
author = "Yunxiang Zhu and Drake, {Matthew M} and Stuart Kornfeld",
year = "1999",
month = "4",
day = "27",
doi = "10.1073/pnas.96.9.5013",
language = "English (US)",
volume = "96",
pages = "5013--5018",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "9",

}

TY - JOUR

T1 - ADP-ribosylation factor 1 dependent clathrin-coat assembly on synthetic liposomes

AU - Zhu, Yunxiang

AU - Drake, Matthew M

AU - Kornfeld, Stuart

PY - 1999/4/27

Y1 - 1999/4/27

N2 - The assembly of clathrin-coated vesicles on Golgi membranes is initiated by the GTP-binding protein ADP ribosylation factor (ARF), which generates high-affinity membrane-binding sites for the heterotetrameric AP-1 adaptor complex. Once bound, the AP-1 recruits clathrin triskelia, which polymerize to form the coat. We have found that ARF·GTP also recruits AP-1 and clathrin onto protein-free liposomes. The efficiency of this process is modulated by the composition of the liposomes, with phosphatidylserine being the most stimulatory phospholipid. There is also a requirement for cytosolic factor(s) other than ARF. Thin-section electron microscopy shows the presence of clathrin-coated buds and vesicles that resemble those formed in vivo. These results indicate that AP-1-containing clathrin-coated vesicles can form in the absence of integral membrane proteins. Thus, ARF·GTP, appropriate lipids, and cytosolic factor(s) are the minimal components necessary for AP- 1 clathrin-coat assembly.

AB - The assembly of clathrin-coated vesicles on Golgi membranes is initiated by the GTP-binding protein ADP ribosylation factor (ARF), which generates high-affinity membrane-binding sites for the heterotetrameric AP-1 adaptor complex. Once bound, the AP-1 recruits clathrin triskelia, which polymerize to form the coat. We have found that ARF·GTP also recruits AP-1 and clathrin onto protein-free liposomes. The efficiency of this process is modulated by the composition of the liposomes, with phosphatidylserine being the most stimulatory phospholipid. There is also a requirement for cytosolic factor(s) other than ARF. Thin-section electron microscopy shows the presence of clathrin-coated buds and vesicles that resemble those formed in vivo. These results indicate that AP-1-containing clathrin-coated vesicles can form in the absence of integral membrane proteins. Thus, ARF·GTP, appropriate lipids, and cytosolic factor(s) are the minimal components necessary for AP- 1 clathrin-coat assembly.

KW - AP-1 adaptor complex

UR - http://www.scopus.com/inward/record.url?scp=0033609083&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033609083&partnerID=8YFLogxK

U2 - 10.1073/pnas.96.9.5013

DO - 10.1073/pnas.96.9.5013

M3 - Article

VL - 96

SP - 5013

EP - 5018

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 9

ER -