A requirement for ARF6 in Fcγ receptor-mediated phagocytosis in macrophages

Qing Zhang, Dianne Cox, Ching Chun Tseng, Julie G. Donaldson, Steven Greenberg

Research output: Contribution to journalArticle

166 Citations (Scopus)

Abstract

Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcγ receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The addition of 100 μM BFA to a subclone of RAW 264.7 macrophages disrupted the appearance and function of the Golgi apparatus as indicated by altered immunofluorescent distribution of β-COP and reduced efflux of BODIPY C5-ceramide, a phospholipid that normally accumulates in the Golgi apparatus. In contrast, BFA had no effect on phagocytosis of IgG-coated erythrocytes. These results suggested that activation of BFA-sensitive ARFs is not required for phagocytosis. ARF6 is unique among members of the ARF family in that its membrane association is unaffected by BFA. Expression of ARF6 mutants defective in either GTP hydrolysis (Q67L) or binding (T27N) inhibited phagocytosis of IgG-coated erythrocytes and attenuated the focal accumulation of F-actin beneath the test particles. These results indicate a requirement for ARF6 in Fcγ receptor-mediated phagocytosis and suggest that ARF6 is an important mediator of cytoskeletal alterations after Fcγ receptor activation.

Original languageEnglish (US)
Pages (from-to)19977-19981
Number of pages5
JournalJournal of Biological Chemistry
Volume273
Issue number32
DOIs
StatePublished - Aug 7 1998
Externally publishedYes

Fingerprint

Brefeldin A
Fc Receptors
Macrophages
Phagocytosis
Membranes
Golgi Apparatus
Actins
Immunoglobulin G
Chemical activation
Erythrocytes
Ceramides
GTP Phosphohydrolases
Metabolites
Guanosine Triphosphate
Membrane Fusion
Pseudopodia
Hydrolysis
Phospholipids
Fusion reactions
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

A requirement for ARF6 in Fcγ receptor-mediated phagocytosis in macrophages. / Zhang, Qing; Cox, Dianne; Tseng, Ching Chun; Donaldson, Julie G.; Greenberg, Steven.

In: Journal of Biological Chemistry, Vol. 273, No. 32, 07.08.1998, p. 19977-19981.

Research output: Contribution to journalArticle

Zhang, Qing ; Cox, Dianne ; Tseng, Ching Chun ; Donaldson, Julie G. ; Greenberg, Steven. / A requirement for ARF6 in Fcγ receptor-mediated phagocytosis in macrophages. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 32. pp. 19977-19981.
@article{39abc5ea8a70494fa32117c02b972337,
title = "A requirement for ARF6 in Fcγ receptor-mediated phagocytosis in macrophages",
abstract = "Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcγ receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The addition of 100 μM BFA to a subclone of RAW 264.7 macrophages disrupted the appearance and function of the Golgi apparatus as indicated by altered immunofluorescent distribution of β-COP and reduced efflux of BODIPY C5-ceramide, a phospholipid that normally accumulates in the Golgi apparatus. In contrast, BFA had no effect on phagocytosis of IgG-coated erythrocytes. These results suggested that activation of BFA-sensitive ARFs is not required for phagocytosis. ARF6 is unique among members of the ARF family in that its membrane association is unaffected by BFA. Expression of ARF6 mutants defective in either GTP hydrolysis (Q67L) or binding (T27N) inhibited phagocytosis of IgG-coated erythrocytes and attenuated the focal accumulation of F-actin beneath the test particles. These results indicate a requirement for ARF6 in Fcγ receptor-mediated phagocytosis and suggest that ARF6 is an important mediator of cytoskeletal alterations after Fcγ receptor activation.",
author = "Qing Zhang and Dianne Cox and Tseng, {Ching Chun} and Donaldson, {Julie G.} and Steven Greenberg",
year = "1998",
month = "8",
day = "7",
doi = "10.1074/jbc.273.32.19977",
language = "English (US)",
volume = "273",
pages = "19977--19981",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "32",

}

TY - JOUR

T1 - A requirement for ARF6 in Fcγ receptor-mediated phagocytosis in macrophages

AU - Zhang, Qing

AU - Cox, Dianne

AU - Tseng, Ching Chun

AU - Donaldson, Julie G.

AU - Greenberg, Steven

PY - 1998/8/7

Y1 - 1998/8/7

N2 - Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcγ receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The addition of 100 μM BFA to a subclone of RAW 264.7 macrophages disrupted the appearance and function of the Golgi apparatus as indicated by altered immunofluorescent distribution of β-COP and reduced efflux of BODIPY C5-ceramide, a phospholipid that normally accumulates in the Golgi apparatus. In contrast, BFA had no effect on phagocytosis of IgG-coated erythrocytes. These results suggested that activation of BFA-sensitive ARFs is not required for phagocytosis. ARF6 is unique among members of the ARF family in that its membrane association is unaffected by BFA. Expression of ARF6 mutants defective in either GTP hydrolysis (Q67L) or binding (T27N) inhibited phagocytosis of IgG-coated erythrocytes and attenuated the focal accumulation of F-actin beneath the test particles. These results indicate a requirement for ARF6 in Fcγ receptor-mediated phagocytosis and suggest that ARF6 is an important mediator of cytoskeletal alterations after Fcγ receptor activation.

AB - Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcγ receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The addition of 100 μM BFA to a subclone of RAW 264.7 macrophages disrupted the appearance and function of the Golgi apparatus as indicated by altered immunofluorescent distribution of β-COP and reduced efflux of BODIPY C5-ceramide, a phospholipid that normally accumulates in the Golgi apparatus. In contrast, BFA had no effect on phagocytosis of IgG-coated erythrocytes. These results suggested that activation of BFA-sensitive ARFs is not required for phagocytosis. ARF6 is unique among members of the ARF family in that its membrane association is unaffected by BFA. Expression of ARF6 mutants defective in either GTP hydrolysis (Q67L) or binding (T27N) inhibited phagocytosis of IgG-coated erythrocytes and attenuated the focal accumulation of F-actin beneath the test particles. These results indicate a requirement for ARF6 in Fcγ receptor-mediated phagocytosis and suggest that ARF6 is an important mediator of cytoskeletal alterations after Fcγ receptor activation.

UR - http://www.scopus.com/inward/record.url?scp=0032493491&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032493491&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.32.19977

DO - 10.1074/jbc.273.32.19977

M3 - Article

VL - 273

SP - 19977

EP - 19981

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 32

ER -