Targeting of Protein Kinase C-ϵ during Fcγ Receptor-dependent Phagocytosis Requires the ϵC1B Domain and Phospholipase C-γ1

Cheeseman, Keylon; Ueyama, Takeshi; Michaud, Tanya M.; Kashiwagi, Kaori; Wang, Demin; Flax, Lindsay A.; Shirai, Yasuhito; Loegering, Daniel J.; Saito, Naoaki; Lennartz, Michelle R.

doi:10.1091/mbc.e04-12-1100

Cited by 50 publications

(74 citation statements)

References 48 publications

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“…Because PLC␥2 is expressed at higher levels than PLC␥1 in RAW cells (19), we investigated the role of Tec family kinases in PLC␥2 activation in these cells stimulated by IgG-RBC (Fig. 6).…”

Section: Requirement For Btk and Tec In Downstream Signaling During Lmentioning

confidence: 99%

“…DAG can be generated through the activities of phospholipase D or PLC, and there is support for the involvement of both enzymes in phagocytosis (15)(16)(17). Recently, PLC␥-derived DAG was shown to be important for the activation of protein kinase C (PKC), which accumulates at phagocytic cups and plays a critical role in Fc␥R-mediated phagocytosis (18,19).…”

mentioning

confidence: 99%

“…PLC␥2 clustered at phagocytic cups may be part of such complexes. However, PLC␥2 involvement in Fc␥R-mediated phagocytosis has recently been questioned (19). This is primarily because phagocytosis through Fc␥R was not significantly affected by PLC␥2 deficiency in macrophages from PLC␥2 Ϫ/Ϫ mice (19,31).…”

mentioning

confidence: 99%

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Dual Functions of Bruton’s Tyrosine Kinase and Tec Kinase during Fcγ Receptor-Induced Signaling and Phagocytosis

Jongstra‐Bilen

Cano

Hasija

et al. 2008

The Journal of Immunology

121

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Tec family nonreceptor tyrosine kinases are expressed by hematopoietic cells, activate phospholipase C (PLC)γ, and regulate cytoskeletal rearrangement, yet their role in FcγR-induced signaling and phagocytosis remains unknown. We demonstrate in this study that Bruton’s tyrosine kinase (Btk) and Tec, the only Tec kinases expressed by RAW 264.7 cells, are activated throughout phagocytosis. Activated Btk and Tec kinase accumulate at an early stage at the base of phagocytic cups and inhibition of their activity by the specific inhibitor LFM-A13 or expression by small interfering RNA significantly inhibited FcγR-induced phagocytosis. Similarly, a significant role for these kinases in phagocytosis was found in primary macrophages. FcγR-induced activation of Mac-1, which is required for optimal phagocytosis, was markedly inhibited and our findings suggest that the roles of kinases Btk and Tec in Mac-1 activation account for their functions in the early stages of phagocytosis. Initial activation of PLCγ2, the predominant PLC isoform in RAW 264.7 cells, is dependent on Syk. In contrast, a late and prolonged activation of PLCγ2 was dependent on Btk and Tec. We found accumulation of diacylglycerol (DAG), a PLCγ product, in phagosome membranes, and activated Btk, but not Tec, colocalized with phagosomal DAG. Inhibition of Tec family kinase activity increased the level of DAG in phagosomes, suggesting a negative regulatory role for Btk. Tec, in contrast, clustered at sites near phagosome formation. In summary, we elucidated that Tec family kinases participate in at least two stages of FcγR-mediated phagocytosis: activation of Mac-1 during ingestion, and after phagosome formation, during which Btk and Tec potentially have distinct roles.

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Section: Requirement For Btk and Tec In Downstream Signaling During Lmentioning

confidence: 99%

mentioning

confidence: 99%

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Dual Functions of Bruton’s Tyrosine Kinase and Tec Kinase during Fcγ Receptor-Induced Signaling and Phagocytosis

Jongstra‐Bilen

Cano

Hasija

et al. 2008

The Journal of Immunology

121

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“…First, antagonists of PLC severely impair phagocytosis by macrophages (7,12). This inhibition is not mimicked by preventing the associated [Ca 2ϩ ] transient, suggesting that DAG, and not inositol 1,4,5-trisphosphate, is the crucial product of the PLC (13).…”

mentioning

confidence: 99%

“…Proteins bearing C1 domains include, most notably, members of the classical and novel families of protein kinase C (PKC), making them suitable candidates to account for the DAG dependence of phagocytosis. Indeed, PKC␣, a classical isoform, and PKC⑀ and PKC␦, both novel isoforms, are recruited to phagosomes (12,15,17,18). Although the role of the various PKC isoforms in particle engulfment has been equivocal over the years, Cheeseman et al (12) convincingly demonstrated that PKC⑀ contributes to particle uptake in a PLC-and DAG-dependent manner.…”

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confidence: 99%

Localized Diacylglycerol-dependent Stimulation of Ras and Rap1 during Phagocytosis

Botelho

Harrison

Stone

et al. 2009

Journal of Biological Chemistry

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We describe a role for diacylglycerol in the activation of Ras and Rap1 at the phagosomal membrane. During phagocytosis, Ras density was similar on the surface and invaginating areas of the membrane, but activation was detectable only in the latter and in sealed phagosomes. Ras activation was associated with the recruitment of RasGRP3, a diacylglycerol-dependent Ras/ Rap1 exchange factor. Recruitment to phagosomes of RasGRP3, which contains a C1 domain, parallels and appears to be due to the formation of diacylglycerol. Accordingly, Ras and Rap1 activation was precluded by antagonists of phospholipase C and of diacylglycerol binding. Ras is dispensable for phagocytosis but controls activation of extracellular signal-regulated kinase, which is partially impeded by diacylglycerol inhibitors. By contrast, cross-activation of complement receptors by stimulation of Fc␥ receptors requires Rap1 and involves diacylglycerol. We suggest a role for diacylglycerol-dependent exchange factors in the activation of Ras and Rap1, which govern distinct processes induced by Fc␥ receptor-mediated phagocytosis to enhance the innate immune response.Receptors that interact with the constant region of IgG (Fc␥R) 4 mediate the recognition and elimination of soluble immune complexes and particles coated (opsonized) with immunoglobulins. Clustering of Fc␥R on the surface of leukocytes upon attachment to multivalent ligands induces their activation and subsequent internalization. Soluble immune complexes are internalized by endocytosis, a clathrin-and ubiquitylation-dependent process (1). In contrast, large, particulate complexes like IgG-coated pathogens are ingested by phagocytosis, a process that is contingent on extensive actin polymerization that drives the extension of pseudopods (2). In parallel with the internalization of the opsonized targets, crosslinking of phagocytic receptors triggers a variety of other responses that are essential components of the innate immune response. These include degranulation, activation of the respiratory burst, and the synthesis and release of multiple inflammatory agents (3, 4).Like T and B cell receptors, Fc␥R possesses an immunoreceptor tyrosine-based activation motif that is critical for signal transduction (3, 4). Upon receptor clustering, tyrosyl residues of the immunoreceptor tyrosine-based activation motif are phosphorylated by Src family kinases, thereby generating a docking site for Syk, a tyrosine kinase of the ZAP70 family (3, 4). The recruitment and activation of Syk in turn initiates a cascade of events that include activation of Tec family kinases, Rho-and ARF-family GTPases, phosphatidylinositol 3-kinase, phospholipase C␥ (PLC␥), and a multitude of additional effectors that together remodel the underlying cytoskeleton, culminating in internalization of the bound particle (5, 6).Phosphoinositide metabolism is thought to be critical for Fc␥R-induced phagocytosis (7,8). Highly localized and very dynamic phosphoinositide changes have been observed at sites of phagocytosis: phosphatidyli...

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Macrophage ICAM-1 functions as a regulator of phagocytosis in LPS induced endotoxemia

et al. 2021

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Objective Intracellular adhesion molecule-1 (ICAM-1), a transmembrane glycoprotein belonging to the immunoglobulin superfamily, plays a critical role in mediating cell-cell interaction and outside-in cell signaling during the immune response. ICAM-1 is expressed on the cell surface of several cell types including endothelial cells, epithelial cells, leucocytes, fibroblasts, and neutrophils. Despite ICAM-1 has been detected on macrophage, little is known about the function and mechanism of macrophage ICAM-1. Methods To investigate the role of lipopolysaccharide (LPS) in ICAM-1 regulation, both the protein and cell surface expression of ICAM-1 were measured. The phagocytosis of macrophage was evaluated by flow cytometry and Confocal microscopy. Small interfering RNA and neutralizing antibody of ICAM-1 were used to assess the effect of ICAM-1 on macrophage phagocytosis. TLR4 gene knockout mouse and cytoplasmic and mitochondrial ROS scavenger were used for the regulation of ICAM-1 expression. ROS was determined using flow cytometry. Results In this study, we reported that macrophage can be stimulated to increase both the protein and cell surface expression of ICAM-1 by LPS. Macrophage ICAM-1 expression was correlated with enhanced macrophage phagocytosis. We found that using ICAM-1 neutralizing antibody or ICAM-1 silencing to attenuate the function or expression of ICAM-1 could decrease LPS-induced macrophage phagocytosis. Furthermore, we found that knocking out of TLR4 led to inhibited cytoplasmic and mitochondrial ROS production, which in turn, attenuated ICAM-1 expression at both the protein and cell surface levels. Conclusion This study demonstrates that the mechanism of ICAM-1-mediated macrophage phagocytosis is depending on TLR4-mediated ROS production and provides significant light on macrophage ICAM-1 in endotoxemia.

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Targeting of Protein Kinase C-ϵ during Fcγ Receptor-dependent Phagocytosis Requires the ϵC1B Domain and Phospholipase C-γ1

Cited by 50 publications

References 48 publications

Dual Functions of Bruton’s Tyrosine Kinase and Tec Kinase during Fcγ Receptor-Induced Signaling and Phagocytosis

Dual Functions of Bruton’s Tyrosine Kinase and Tec Kinase during Fcγ Receptor-Induced Signaling and Phagocytosis

Localized Diacylglycerol-dependent Stimulation of Ras and Rap1 during Phagocytosis

Macrophage ICAM-1 functions as a regulator of phagocytosis in LPS induced endotoxemia

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