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Cited by 166Protein Kinase Regulation and GTPase SignalingPhagocytosis and Immune RegulationCellular transport and secretion

Rho is required for the initiation of calcium signaling and phagocytosis by Fcγ receptors in macrophages

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Hackam, David J., Rotstein, Ori D., Schreiber, Alan, Zhang, Wei-jian, Grinstein, Sergio·1997·The Journal of Experimental Medicine
DOI 10.1084/jem.186.6.955PMC PMC2199044
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Abstract

Phagocytosis of bacteria by macrophages and neutrophils is an essential component of host defense against infection. The mechanism whereby the interaction of opsonized particles with Fcγ receptors triggers the engulfment of opsonized particles remains incompletely understood, although activation of tyrosine kinases has been recognized as an early step. Recent studies in other systems have demonstrated that tyrosine kinases can in turn signal the activation of small GTPases of the ras superfamily. We therefore investigated the possible role of Rho in Fc receptor–mediated phagocytosis. To this end we microinjected J774 macrophages with C3 exotoxin from Clostridium botulinum, which ADP-ribosylates and inactivates Rho. C3 exotoxin induced the retraction of filopodia, the disappearance of focal complexes, and a global decrease in the F-actin content of J774 cells. In addition, these cells exhibited increased spreading and the formation of vacuolar structures. Importantly, inactivation of Rho resulted in the complete abrogation of phagocytosis. Inhibition of Fcγ receptor–mediated phagocytosis by C3 exotoxin was confirmed in COS cells, which become phagocytic upon transfection of the FcγRIIA receptor. Rho was found to be essential for the accumulation of phosphotyrosine and of F-actin around phagocytic cups and for Fcγ receptor–mediated Ca2+ signaling. The clustering of receptors in response to opsonin, an essential step in Fcγ-induced signaling, was the earliest event shown to be inhibited by C3 exotoxin. The effect of the toxin was specific, since clustering and internalization of transferrin receptors were unaffected by microinjection of C3. These data identify a role for small GTPases in Fcγ receptor–mediated phagocytosis by leukocytes.

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  • Actin assembly during phagocytosis is mediated by Rho family GTPases.

    “Experiments using clostridial toxins or transfection of dominant-negative constructs have convincingly demonstrated that actin assembly during phagocytosis is mediated by Rho family GTPases (Cox et al. 1997; Hackam et al. 1997; Caron and Hall 1998; Massol et al. 1998).”

    From Localized Biphasic Changes in Phosphatidylinositol-4,5-Bisphosphate at Sites of Phagocytosis (2000) · cites this paper as Hackam et al. 1997

    The Results report that inactivating Rho eliminated F-actin cups during Fcγ receptor–mediated phagocytosis, supporting Rho's role in actin assembly. The Discussion confines the conclusions to transformed cell lines; this paper alone does not establish the claim for phagocytosis generally. (LLM, conf=0.85)

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Cited by

Sorted by citations

Authors
Hackam, D. J., Rotstein, O. D., Schreiber, A., Zhang, W.-J., & Grinstein, S.
Affiliations
Hospital for Sick Children (CA)
Corresponding author
Grinstein, Sergio
ORCID
Venue
The Journal of Experimental Medicine
Publisher
Rockefeller University Press
Published
1997-09-15
Volume
186
Issue
6
Pages
955-966
ISSN
  • 0022-1007 (ISSN-L)
  • 1540-9538
DOI
10.1084/jem.186.6.955
PMC
PMC2199044
OpenAlex
W2023497277
Cited by
166
References
1
Citations by year
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FWCI
6.77
Type
journal article
Language
en
Open access
Locations
  • The Journal of Experimental Medicine (publishedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
  • Howard Hughes Medical Institute
  • Medical Research Council Canada
  • Medical Research Council
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Immunology and Microbiology › Immunology › Phagocytosis and Immune Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
Keywords
Rho GTPases, Fc gamma receptor, phagocytosis, calcium signaling, macrophages, small GTPases, FCGR2A, receptor clustering, F-actin, filopodia, tyrosine phosphorylation, opsonization, phagocytic cup
MeSH terms
Actins, Animals, Botulinum Toxins, Calcium, Cell Line, Exotoxins, Hydrogen-Ion Concentration, Macrophages, Membrane Proteins, Phagocytosis, Transfection, Signal Transduction, Receptors, IgG, Phosphotyrosine, GTP-Binding Proteins, COS Cells, rhoB GTP-Binding Protein, Mice

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