A role for PKC-epsilon in Fc gamma R-mediated phagocytosis by RAW 264.7 cells
(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:
https://lup.lub.lu.se/record/892132
- author
- Larsen, EC ; Ueyama, T ; Brannock, PM ; Shirai, Y ; Saito, N ; Larsson, Christer LU ; Loegering, D ; Weber, PB and Lennartz, MR
- organization
- publishing date
- 2002
- 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
- pmid:12499353
- ISSN
- 0021-9525
- DOI
- 10.1083/jcb.200205140
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Tumour Cell Biology (013017530)
- 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
- 2016-04-01 11:59:22
- date last changed
- 2022-01-26 21:11:45
@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}}, keywords = {{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}}, doi = {{10.1083/jcb.200205140}}, volume = {{159}}, year = {{2002}}, }