2026-07-22T00:21:31Zhttps://repisalud.isciii.es/rest/oai/request

oai:repisalud.isciii.es:20.500.12105/51242024-09-27T08:17:01Zcom_20.500.12105_19604com_20.500.12105_2051col_20.500.12105_19605col_20.500.12105_19607

00925njm 22002777a 4500 dc Garcia-Ortiz, Almudena author Martin-Cofreces, Noa B. author Ibiza, Sales author Ortega, Angel author Izquierdo-Alvarez, Alicia author Trullo, Antonio author Victor, Victor Manuel author Calvo, Enrique author Sot, Begona author Martinez-Ruiz, Antonio author Vazquez, Jesus author Sanchez-Madrid, Francisco author Serrador, Juan M. author 2017 The actin cytoskeleton coordinates the organization of signaling microclusters at the immune synapse (IS); however, the mechanisms involved remain poorly understood. We show here that nitric oxide (NO) generated by endothelial nitric oxide synthase (eNOS) controls the coalescence of protein kinase C-theta (PKC-theta) at the central supramolecular activation cluster (c-SMAC) of the IS. eNOS translocated with the Golgi to the IS and partially colocalized with F-actin around the c-SMAC. This resulted in reduced actin polymerization and centripetal retrograde flow of beta-actin and PKC-theta from the lamellipodium-like distal (d)-SMAC, promoting PKC-theta. activation. Furthermore, eNOS-derived NO S-nitrosylated beta-actin on Cys374 and impaired actin binding to profilin-1 (PFN1), as confirmed with the transnitrosylating agent S-nitroso-L-cysteine (Cys-NO). The importance of NO and the formation of PFN1-actin complexes on the regulation of PKC-theta. was corroborated by overexpression of PFN1- and actin-binding defective mutants of beta-actin (C374S) and PFN1 (H119E), respectively, which reduced the coalescence of PKC-theta. at the c-SMAC. These findings unveil a novel NO-dependent mechanism by which the actin cytoskeleton controls the organization and activation of signaling microclusters at the IS. PLoS Biol. 2017; 15(4):e2000653 10.1371/journal.pbio.2000653 1545-7885 Plos Biology 28394935 http://hdl.handle.net/20.500.12105/5124 NITRIC-OXIDE SYNTHASE T-CELL-ACTIVATION IMMUNOLOGICAL SYNAPSE RETROGRADE FLOW ARP2/3 COMPLEX PROTEIN RECEPTOR POLYMERIZATION POLARIZATION CYTOSKELETON eNOS S-nitrosylates beta-actin on Cys374 and regulates PKC-theta at the immune synapse by impairing actin binding to profilin-1