Endosomal clathrin drives actin accumulation at the immunological synapse

Calabia-Linares, Carmen; Robles‐Valero, Javier; Fuente, Hortensia de la; Pérez‐Martínez, Manuel; Martín-Cófreces, Noa B.; Alfonso-Pérez, Manuel; Gutiérrez-Vázquez, Cristina; Mittelbrunn, Marı́a; Ibiza, Sales; Urbano-Olmos, Francisco R.; Aguado-Ballano, C; Sánchez-Sorzano, Carlos Óscar; Sánchez‐Madrid, Francisco; Veiga, Esteban

doi:10.1242/jcs.078832

Cited by 79 publications

(79 citation statements)

References 53 publications

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“…Yeast with similar properties function with only one CLC, which is most homologous to CLCa and is a key player in endocytosis through its association with the actin-binding Hip homolog Sla2p, which suggests that CLCa could confer a specialized actin-interacting activity that is critical for clathrin function in lymphocytes. Indeed, both lymphocyte endocytosis and immune synapse formation depend on cooperation between clathrin and actin (38,39). An alternative possible explanation for the conservation of CLC isoforms is that CLCa and CLCb influence cargo selection differently, which could also have tissue-specific and physiological consequences.…”

Section: Clc Depletion From Cell Lines Establishes Selectivity For DImentioning

confidence: 99%

Clathrin light chains’ role in selective endocytosis influences antibody isotype switching

Majeed

Evans

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Clathrin, a cytosolic protein composed of heavy and light chain subunits, assembles into a vesicle coat, controlling receptor-mediated endocytosis. To establish clathrin light chain (CLC) function in vivo, we engineered mice lacking CLCa, the major CLC isoform in B lymphocytes, generating animals with CLC-deficient B cells. In CLCa-null mice, the germinal centers have fewer B cells, and they are enriched for IgA-producing cells. This enhanced switch to IgA production in the absence of CLCa was attributable to increased transforming growth factor β receptor 2 (TGFβR2) signaling resulting from defective endocytosis. Internalization of C-X-C chemokine receptor 4 (CXCR4), but not CXCR5, was affected in CLCa-null B cells, and CLC depletion from cell lines affected endocytosis of the δ-opioid receptor, but not the β2-adrenergic receptor, defining a role for CLCs in the uptake of a subset of signaling receptors. This instance of clathrin subunit deletion in vertebrates demonstrates that CLCs contribute to clathrin’s role in vivo by influencing cargo selectivity, a function previously assigned exclusively to adaptor molecules.

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Section: Clc Depletion From Cell Lines Establishes Selectivity For DImentioning

confidence: 99%

Clathrin light chains’ role in selective endocytosis influences antibody isotype switching

Majeed

Evans

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…Protein relocalization to the IS was quantified using the Synapsemeasures plugin in Image J [26]. Briefly, the quantification takes into account the fluorescence intensity signals measured in selected regions with similar areas at the T-APC contact zone (IS), the APC membrane not in contact with the T cell (B), the T cell membrane not in contact with the APC (T), and the background (Bg).…”

Section: Quantitative Imaging Analysesmentioning

confidence: 99%

Tetraspanins CD9 and CD151 at the immune synapse support T‐cell integrin signaling

et al. 2014

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Understanding how the immune response is activated and amplified requires detailed knowledge of the stages in the formation of the immunological synapse (IS) between T lymphocytes and antigen-presenting cells (APCs). We show that tetraspanins CD9 and CD151 congregate at the T-cell side of the IS. Silencing of CD9 or CD151 blunts the IL-2 secretion and expression of the activation marker CD69 by APC-conjugated T lymphocytes, but does not affect the accumulation of CD3 or actin to the IS, or the translocation of the microtubule-organizing center toward the T-B contact area. CD9 or CD151 silencing diminishes the relocalization of α4β1 integrin to the IS and reduces the accumulation of high-affinity β1 integrins at the cell-cell contact. These changes are accompanied by diminished phosphorylation of the integrin downstream targets FAK and ERK1/2. Our results suggest that CD9 and CD151 support integrin-mediated signaling at the IS.Keywords: CD9 r CD151 r Immune synapse r Integrins r T-cell activation Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionInteraction between T lymphocytes and antigen-presenting cells (APCs) is critical for antigen presentation and for the initiation of the immune response. This process is characterized by the formation of a dynamic structure called the immune synapse (IS) at the contact between T cells and APCs [1]. At the T cell side of the IS, T-cell receptors (TCRs) and Correspondence: Dr.Francisco Sánchez-Madrid e-mail: fsmadrid@salud.madrid.org associated molecules accumulate in the central area (central supramolecular activation cluster; cSMAC), while adhesion receptors and integrins reorganize in a surrounding external ring called peripheral SMAC (pSMAC) [1]. The increased IS stability enhances T-cell sensitivity, stimulates cytoskeletal processes, induces downstream signaling, and drives effective T-cell differentiation [2,3]. How integrins regulate these processes is not fully understood.Several membrane receptors and integrins concentrated at the IS -CD3, CD4, ICAM-1, lymphocyte function-associated antigen 1 (LFA-1), and very late antigen 4 (VLA-4) -are associated with tetraspanins [4,5] 1968 Rocha-Perugini Vera et al. Eur. J. Immunol. 2014. 44: 1967-1975 organize membrane macrocomplexes called tetraspanin-enriched microdomains (TEMs) [6]. Tetraspanins modulate the function of their associated partners and play important roles in immunity, inflammation, and other processes [6]. Tetraspanins that accumulate at the IS include CD81, which regulates IS architectural organization and maturation [5], and CD82, which contributes to T-cell activation through Rho GTPase-dependent regulation of the actin cytoskeleton [7]. CD82 also increases the adhesion of LFA-1 integrin to its ligand ICAM-1 expressed on APCs [4]. The tetraspanins CD81, CD82, CD53, and CD9 provide T cell costimulatory signals, and CD9 localizes together with TCR signaling molecules in lipid microdomains [4]. Moreover, the T cells of mice deficien...

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“…For F-actin and profilin accumulation at intercellular contacts, the Image J plug-in "Synapsemeasure" was used as described previously (45). Briefly, by selecting regions of interest of the same area for all measurements, the fluorescence intensity at the cell to cell contact area (V), an area of the infected cell not in contact with the target cell (I), an area of the target T cell not in contact with the infected cell (T), and the background (Bg) were quantified.…”

Section: Methodsmentioning

confidence: 99%

Actin-binding Protein Drebrin Regulates HIV-1-triggered Actin Polymerization and Viral Infection

Gordon-Alonso

Rocha-Perugini

Álvarez

et al. 2013

Journal of Biological Chemistry

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Background: Drebrin binds to F-actin and CXCR4 in T cells. Thus, it is a potential candidate for the modulation of HIV-1 infection. Results: Drebrin and CXCR4 accumulate at viral attachment areas. Drebrin knockdown decreases F-actin polymerization, and increases local profilin accumulation and HIV-1 infection. Conclusion: Drebrin inhibits HIV-1 entry by stabilizing HIV-1-triggered F-actin polymerization. Significance: Modulation of actin dynamics differentially regulates each viral step for an effective viral infection. HIV-1 contact with target cells triggers F-actin rearrangements that are essential for several steps of the viral cycle. Successful HIV entry into CD4؉ T cells requires actin reorganization induced by the interaction of the cellular receptor/co-receptor complex CD4/CXCR4 with the viral envelope complex gp120/gp41 (Env).In this report, we analyze the role of the actin modulator drebrin in HIV-1 viral infection and cell to cell fusion. We show that drebrin associates with CXCR4 before and during HIV infection. Drebrin is actively recruited toward cellvirus and Env-driven cell to cell contacts. After viral internalization, drebrin clustering is retained in a fraction of the internalized particles. Through a combination of RNAi-based inhibition of endogenous drebrin and GFP-tagged expression of wild-type and mutant forms, we establish drebrin as a negative regulator of HIV entry and HIV-mediated cell fusion. Down-regulation of drebrin expression promotes HIV-1 entry, decreases F-actin polymerization, and enhances profilin local accumulation in response to HIV-1. These data underscore the negative role of drebrin in HIV infection by modulating viral entry, mainly through the control of actin cytoskeleton polymerization in response to HIV-1.Human immunodeficiency virus (HIV)-1 entry requires fusion between the viral envelope and the plasma membrane of the target cell (1). This process is mediated by the viral envelope glycoprotein complex gp120/gp41 (Env), 5 which interacts first with CD4 and then with a co-receptor, one of the chemokine receptors CCR5 or CXCR4 (2, 3).For the viral and cellular membranes to fuse, a critical number of gp120-CD4/coreceptor engagements are needed to establish an energetically productive fusion pore (4). HIV-1 can also be transmitted from infected to uninfected cells through cell to cell contacts, called virological synapses because of their similarities to the immunological synapse (5). As an example, CD4 and CXCR4 recruitment (6 -8) and local actin polymerization take place in both processes (6, 9 -11). Receptor clustering is regulated by two main factors: 1) insertion into specific membrane domains by lateral membrane receptor interactions (12) such as lipid rafts (13) or tetraspanin-enriched microdomains (14 -17); and 2) actin remodeling (6, 18 -20). This phenomenon is well illustrated by experiments in which HIV-1 contact induces CD4 and CXCR4 capping at the plasma membrane of target CD4 ϩ T lymphocytes (21,22). Receptor capping requires the formation of a subcortical...

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Endosomal clathrin drives actin accumulation at the immunological synapse

Cited by 79 publications

References 53 publications

Clathrin light chains’ role in selective endocytosis influences antibody isotype switching

Clathrin light chains’ role in selective endocytosis influences antibody isotype switching

Tetraspanins CD9 and CD151 at the immune synapse support T‐cell integrin signaling

Actin-binding Protein Drebrin Regulates HIV-1-triggered Actin Polymerization and Viral Infection

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