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A role for PKC-epsilon in Fc gamma R-mediated phagocytosis by RAW 264.7 cells

Larsen, EC; Ueyama, T; Brannock, PM; Shirai, Y; Saito, N; Larsson, Christer LU ; Loegering, D; Weber, PB and Lennartz, MR (2002) In Journal of Cell Biology 159(6). p.939-944
Abstract
Protein kinase C (PKC) plays a prominent role in immune signaling, and the paradigms for isoform selective signaling are beginning to be elucidated. Real-time microscopy was combined with molecular and biochemical approaches to demonstrate a role for PKC-epsilon in Fcgamma receptor (FcgammaR)-dependent phagocytosis. RAW 264.7 macrophages were transfected with GFP-conjugated PKC isoforms, and GFP movement was followed during phagocytosis of fluorescent IgG-opsonized beads. PKC-epsilon, but not PKC-delta, concentrated around the beads. PKC-epsilon accumulation was transient; apparent as a "flash" on target ingestion. Similarly, endogenous PKC-epsilon was specifically recruited to the nascent phagosomes in a time-dependent manner.... (More)
Protein kinase C (PKC) plays a prominent role in immune signaling, and the paradigms for isoform selective signaling are beginning to be elucidated. Real-time microscopy was combined with molecular and biochemical approaches to demonstrate a role for PKC-epsilon in Fcgamma receptor (FcgammaR)-dependent phagocytosis. RAW 264.7 macrophages were transfected with GFP-conjugated PKC isoforms, and GFP movement was followed during phagocytosis of fluorescent IgG-opsonized beads. PKC-epsilon, but not PKC-delta, concentrated around the beads. PKC-epsilon accumulation was transient; apparent as a "flash" on target ingestion. Similarly, endogenous PKC-epsilon was specifically recruited to the nascent phagosomes in a time-dependent manner. Overexpression of PKC-epsilon, but not PKC-alpha, PKC-delta, or PKC-gamma enhanced bead uptake 1.8-fold. Additionally, the rate of phagocytosis in GFP PKC-epsilon expressors was twice that of cells expressing GFP PKC-delta. Expression of the regulatory domain (ERD) and the first variable region (epsilonV1) of PKC-epsilon inhibited uptake, whereas the corresponding PKC-delta region had no effect. Actin polymerization was enhanced on expression of GFP PKC-epsilon and ERD, but decreased in cells expressing epsilonV1, suggesting that the epsilonRD and epsilonV1 inhibition of phagocytosis is not due to effects on actin polymerization. These results demonstrate a role for PKC-epsilon in FcgammaR-mediated phagocytosis that is independent of its effects on actin assembly. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
immunoglobulin, protein kinase C-epsilon, signal transduction, macrophage, confocal
in
Journal of Cell Biology
volume
159
issue
6
pages
939 - 944
publisher
Rockefeller University Press
external identifiers
  • wos:000180150200004
  • pmid:12499353
  • scopus:0037164567
ISSN
0021-9525
DOI
10.1083/jcb.200205140
language
English
LU publication?
yes
id
94c2c8ff-a214-43ac-804d-8433b42776bd (old id 892132)
alternative location
http://www.jcb.org/cgi/content/abstract/159/6/939
date added to LUP
2008-01-23 13:11:17
date last changed
2017-08-06 03:38:42
@article{94c2c8ff-a214-43ac-804d-8433b42776bd,
  abstract     = {Protein kinase C (PKC) plays a prominent role in immune signaling, and the paradigms for isoform selective signaling are beginning to be elucidated. Real-time microscopy was combined with molecular and biochemical approaches to demonstrate a role for PKC-epsilon in Fcgamma receptor (FcgammaR)-dependent phagocytosis. RAW 264.7 macrophages were transfected with GFP-conjugated PKC isoforms, and GFP movement was followed during phagocytosis of fluorescent IgG-opsonized beads. PKC-epsilon, but not PKC-delta, concentrated around the beads. PKC-epsilon accumulation was transient; apparent as a "flash" on target ingestion. Similarly, endogenous PKC-epsilon was specifically recruited to the nascent phagosomes in a time-dependent manner. Overexpression of PKC-epsilon, but not PKC-alpha, PKC-delta, or PKC-gamma enhanced bead uptake 1.8-fold. Additionally, the rate of phagocytosis in GFP PKC-epsilon expressors was twice that of cells expressing GFP PKC-delta. Expression of the regulatory domain (ERD) and the first variable region (epsilonV1) of PKC-epsilon inhibited uptake, whereas the corresponding PKC-delta region had no effect. Actin polymerization was enhanced on expression of GFP PKC-epsilon and ERD, but decreased in cells expressing epsilonV1, suggesting that the epsilonRD and epsilonV1 inhibition of phagocytosis is not due to effects on actin polymerization. These results demonstrate a role for PKC-epsilon in FcgammaR-mediated phagocytosis that is independent of its effects on actin assembly.},
  author       = {Larsen, EC and Ueyama, T and Brannock, PM and Shirai, Y and Saito, N and Larsson, Christer and Loegering, D and Weber, PB and Lennartz, MR},
  issn         = {0021-9525},
  keyword      = {immunoglobulin,protein kinase C-epsilon,signal transduction,macrophage,confocal},
  language     = {eng},
  number       = {6},
  pages        = {939--944},
  publisher    = {Rockefeller University Press},
  series       = {Journal of Cell Biology},
  title        = {A role for PKC-epsilon in Fc gamma R-mediated phagocytosis by RAW 264.7 cells},
  url          = {http://dx.doi.org/10.1083/jcb.200205140},
  volume       = {159},
  year         = {2002},
}