Differential palmitoylation regulates intracellular patterning of SNAP25

Jennifer Greaves, Luke H Chamberlain

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

SNAP25 regulates membrane fusion events at the plasma membrane and in the endosomal system, and a functional pool of the protein is delivered to recycling endosomes (REs) and the trans Golgi network (TGN) through an ARF6-dependent cycling pathway. SNAP25 is a peripheral membrane protein, and palmitoylation of a cluster of four cysteine residues mediates its stable association with the membrane. Here, we report that palmitoylation also determines the precise intracellular distribution of SNAP25, and that mutating single palmitoylation sites enhances the amount of SNAP25 at the RE and TGN. The farnesylated CAAX motif from Hras was ligated onto a SNAP25 mutant truncated immediately distal to the cysteine-rich domain. This construct displayed the same intracellular distribution as full-length SNAP25, and decreasing the number of cysteine residues in this construct increased its association with the RE and TGN, confirming the dominant role of the cysteine-rich domain in directing the intracellular distribution of SNAP25. Marked differences in the localisations of SNAP25-CAAX and Hras constructs, each with two palmitoylation sites, were observed, showing that subtle differences in palmitoylated sequences can have a major impact upon intracellular targeting. We propose that the cysteine-rich domain of SNAP25 is designed to facilitate the dual function of this SNARE protein at the plasma membrane and endosomes, and that dynamic palmitoylation acts as a mechanism to regulate the precise intracellular patterning of SNAP25.

Original languageEnglish
Pages (from-to)1351-1360
Number of pages10
JournalJournal of Cell Science
Volume124
Issue number8
DOIs
Publication statusPublished - 15 Apr 2011
Externally publishedYes

Fingerprint

Lipoylation
Endosomes
Cysteine
trans-Golgi Network
Cell Membrane
SNARE Proteins
Membrane Fusion
Membrane Proteins
Membranes
Proteins

Keywords

  • Animals
  • Cell Membrane
  • Endosomes
  • Intracellular Space
  • Lipoylation
  • PC12 Cells
  • Protein Transport
  • Rats
  • Synaptosomal-Associated Protein 25
  • trans-Golgi Network
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Differential palmitoylation regulates intracellular patterning of SNAP25. / Greaves, Jennifer; Chamberlain, Luke H.

In: Journal of Cell Science, Vol. 124, No. 8, 15.04.2011, p. 1351-1360.

Research output: Contribution to journalArticle

@article{2df0652fa4834b02ac3cb01c54571a36,
title = "Differential palmitoylation regulates intracellular patterning of SNAP25",
abstract = "SNAP25 regulates membrane fusion events at the plasma membrane and in the endosomal system, and a functional pool of the protein is delivered to recycling endosomes (REs) and the trans Golgi network (TGN) through an ARF6-dependent cycling pathway. SNAP25 is a peripheral membrane protein, and palmitoylation of a cluster of four cysteine residues mediates its stable association with the membrane. Here, we report that palmitoylation also determines the precise intracellular distribution of SNAP25, and that mutating single palmitoylation sites enhances the amount of SNAP25 at the RE and TGN. The farnesylated CAAX motif from Hras was ligated onto a SNAP25 mutant truncated immediately distal to the cysteine-rich domain. This construct displayed the same intracellular distribution as full-length SNAP25, and decreasing the number of cysteine residues in this construct increased its association with the RE and TGN, confirming the dominant role of the cysteine-rich domain in directing the intracellular distribution of SNAP25. Marked differences in the localisations of SNAP25-CAAX and Hras constructs, each with two palmitoylation sites, were observed, showing that subtle differences in palmitoylated sequences can have a major impact upon intracellular targeting. We propose that the cysteine-rich domain of SNAP25 is designed to facilitate the dual function of this SNARE protein at the plasma membrane and endosomes, and that dynamic palmitoylation acts as a mechanism to regulate the precise intracellular patterning of SNAP25.",
keywords = "Animals, Cell Membrane, Endosomes, Intracellular Space, Lipoylation, PC12 Cells, Protein Transport, Rats, Synaptosomal-Associated Protein 25, trans-Golgi Network, Journal Article, Research Support, Non-U.S. Gov't",
author = "Jennifer Greaves and Chamberlain, {Luke H}",
year = "2011",
month = "4",
day = "15",
doi = "10.1242/jcs.079095",
language = "English",
volume = "124",
pages = "1351--1360",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists",
number = "8",

}

TY - JOUR

T1 - Differential palmitoylation regulates intracellular patterning of SNAP25

AU - Greaves, Jennifer

AU - Chamberlain, Luke H

PY - 2011/4/15

Y1 - 2011/4/15

N2 - SNAP25 regulates membrane fusion events at the plasma membrane and in the endosomal system, and a functional pool of the protein is delivered to recycling endosomes (REs) and the trans Golgi network (TGN) through an ARF6-dependent cycling pathway. SNAP25 is a peripheral membrane protein, and palmitoylation of a cluster of four cysteine residues mediates its stable association with the membrane. Here, we report that palmitoylation also determines the precise intracellular distribution of SNAP25, and that mutating single palmitoylation sites enhances the amount of SNAP25 at the RE and TGN. The farnesylated CAAX motif from Hras was ligated onto a SNAP25 mutant truncated immediately distal to the cysteine-rich domain. This construct displayed the same intracellular distribution as full-length SNAP25, and decreasing the number of cysteine residues in this construct increased its association with the RE and TGN, confirming the dominant role of the cysteine-rich domain in directing the intracellular distribution of SNAP25. Marked differences in the localisations of SNAP25-CAAX and Hras constructs, each with two palmitoylation sites, were observed, showing that subtle differences in palmitoylated sequences can have a major impact upon intracellular targeting. We propose that the cysteine-rich domain of SNAP25 is designed to facilitate the dual function of this SNARE protein at the plasma membrane and endosomes, and that dynamic palmitoylation acts as a mechanism to regulate the precise intracellular patterning of SNAP25.

AB - SNAP25 regulates membrane fusion events at the plasma membrane and in the endosomal system, and a functional pool of the protein is delivered to recycling endosomes (REs) and the trans Golgi network (TGN) through an ARF6-dependent cycling pathway. SNAP25 is a peripheral membrane protein, and palmitoylation of a cluster of four cysteine residues mediates its stable association with the membrane. Here, we report that palmitoylation also determines the precise intracellular distribution of SNAP25, and that mutating single palmitoylation sites enhances the amount of SNAP25 at the RE and TGN. The farnesylated CAAX motif from Hras was ligated onto a SNAP25 mutant truncated immediately distal to the cysteine-rich domain. This construct displayed the same intracellular distribution as full-length SNAP25, and decreasing the number of cysteine residues in this construct increased its association with the RE and TGN, confirming the dominant role of the cysteine-rich domain in directing the intracellular distribution of SNAP25. Marked differences in the localisations of SNAP25-CAAX and Hras constructs, each with two palmitoylation sites, were observed, showing that subtle differences in palmitoylated sequences can have a major impact upon intracellular targeting. We propose that the cysteine-rich domain of SNAP25 is designed to facilitate the dual function of this SNARE protein at the plasma membrane and endosomes, and that dynamic palmitoylation acts as a mechanism to regulate the precise intracellular patterning of SNAP25.

KW - Animals

KW - Cell Membrane

KW - Endosomes

KW - Intracellular Space

KW - Lipoylation

KW - PC12 Cells

KW - Protein Transport

KW - Rats

KW - Synaptosomal-Associated Protein 25

KW - trans-Golgi Network

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1242/jcs.079095

DO - 10.1242/jcs.079095

M3 - Article

VL - 124

SP - 1351

EP - 1360

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 8

ER -