The Rho GTPase Rac1 is required for recycling endosome-mediated secretion of TNF in macrophages
Abstract
Rho GTPases are required for many cellular events such as adhesion, motility, and membrane trafficking. Here we show that in macrophages, the Rho GTPases Rac1 and Cdc42 are involved in lamellipodia and filopodia formation, respectively, and that both of these Rho GTPases are essential for the efficient surface delivery of tumor necrosis factor (TNF) to the plasma membrane following TLR4 stimulation. We have previously demonstrated intracellular trafficking of TNF via recycling endosomes in lipopolysaccharide (LPS)‐activated macrophages. Here, we further define a specific role for Rac1 in intracellular TNF trafficking, demonstrating impairment in TNF release following TLR4 stimulation in the presence of a Rac inhibitor, in cells expressing a dominant negative (DN) form of Rac1, and following small interfering RNA (siRNA) knockdown of Rac1. Rac1 activity was required for TNF trafficking but not for TLR4 signaling following LPS stimulation. Reduced TNF secretion was due to a defect in Rac1 activity, but not of the closely related Rho GTPase Rac2, demonstrated by the additional use of macrophages derived from Rac2‐deficient mice. Labeling recycling endosomes by the uptake of fluorescent transferrin enabled us to show that Rac1 was required for the final stages of TNF trafficking and delivery from recycling endosomes to the plasma membrane. Thus, actin remodeling by the Rho GTPase Rac1 is required for TNF cell surface delivery and release from macrophages.
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Rho1 and Cdc42 regulate the post-recycling endosome trafficking of TNF to the plasmalemma.
“Rho1 and Cdc42, two proteins that govern cell shape via actin remodeling, also regulate the post-recycling endosome trafficking of TNF to the plasmalemma (20).”
From Macrophage Cytokines: Involvement in Immunity and Infectious Diseases (2014) · cites this paper as [20]
Cites
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Macrophage Cytokines: Involvement in Immunity and Infectious Diseases
Arango Duque, G., & Descoteaux, A. (2014) · Frontiers in Immunology · doi:10.3389/fimmu.2014.00491 · Cited by 2,708 · see citing works
“Rho1 and Cdc42, two proteins that govern cell shape via actin remodeling, also regulate the postrecycling endosome trafficking of TNF to the plasmalemma (20).”
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“In addition, other molecular players such as Rho GTPases, including Rac1 and Cdc42, play important roles in TNF secretion in macrophage by delivering TNF-carrying recycling endosomes to the cell surface (77).”
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“However, TLR2 pathway causes impairment of TNFa expression visibility at gene level, due to cross-signalling and camouflage (Stanley et al., 2014).”
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“IL-1β is mainly secreted by activated immune cells such as monocytes and macrophages, as well as NK cells, B cells, dendritic cells, fibroblasts, and epithelial cells [8,62].”
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“…were treated with 100 ng ml−1 LPS (to induce TNF synthesis and secretion) and 2 mM of the TACE inhibitor TAPI (to block surface TNF cleavage), for 2 h. Fixed cells were immunostained before and after permeabilization with rabbit and rat TNF antibodies as described previously (Stanley et al., 2014).”
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“Furthermore, it has been shown that TNF a increases the activity of Rac and subsequently activates NADPH oxidases such as Nox1 or Nox2 (82).”
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Deciphering Novel Cytokine Secretion Mechanisms
Lacy, P. (2022) · Scientia · doi:10.33548/scientia843
Self-citationde Freitas, L. G. (2016) · doi:10.35376/10324/18651
“(b) CDC42 induce en los macrófagos la formación de filopodios (57) que son proyecciones citoplasmáticas delgadas que al retraerse alargan la fimbria bacteriana gracias a que se desenrolla la estructura helicoidal cuaternaria de la fimbria.”
Background
- Authors
- Stanley, A. C., Wong, C. X., Micaroni, M., Venturato, J., Khromykh, T., Stow, J. L., & Lacy, P.
- Affiliations
- The University of Queensland (AU)
- Institute of Medical Biology (SG)
- University of Exeter (GB)
- University of Alberta (CA)
- Venue
- Immunology & Cell Biology
- Publisher
- Wiley
- Published
- 2013-12-17
- Volume
- 92
- Issue
- 3
- Pages
- 275-286
- ISSN
- 0004-945X (ISSN-L)
- 0818-9641
- 1440-1711
- OpenAlex
- W1965417957
- Cited by
- 21
- References
- 0
- Citations by year
- 2024: 1 · 2023: 1 · 2022: 2 · 2021: 3 · 2018: 3 · 2017: 2 · 2016: 3 · 2015: 1 · 2014: 5
- FWCI
- 1.74
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Immunology and Cell Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Alberta Heritage Foundation for Medical Research
- Canadian Institutes of Health Research
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Keywords
- Rac1, Rho GTPases, Cdc42, TNF secretion, recycling endosomes, macrophages, TLR4 signaling, lipopolysaccharide, actin cytoskeleton remodeling, lamellipodia, filopodia, Rac2, dominant negative Rac1
- MeSH terms
- RAC2 GTP-Binding Protein, Animals, Cell Line, Cell Membrane, Endocytosis, Macrophages, Mice, Inbred C57BL, Pseudopodia, Pyrones, Quinolines, Endosomes, Recombinant Fusion Proteins, Tumor Necrosis Factor-alpha, Gene Deletion, rac GTP-Binding Proteins, cdc42 GTP-Binding Protein, rac1 GTP-Binding Protein, Protein Transport, RNA, Small Interfering, Green Fluorescent Proteins, Mice, Gene Knockdown Techniques
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