Proteomic Profiling of the TRAF3 Interactome Network Reveals a New Role for the ER-to-Golgi Transport Compartments in Innate Immunity

van Zuylen, Wendy J.; Doyon, Priscilla; Clément, Jean-François; Khan, Kashif Aziz; D'Ambrosio, Lisa M.; Dô, Florence; St-Amant-Verret, Myriam; Wissanji, Tasheen; Emery, Gregory; Gingras, Anne-Claude; Meloche, Sylvain; Servant, Marc J.

Although Bouwmeester and colleagues reported an NF-kBinducing kinase-dependent interaction between Sec16A and NF-kB 2/p100 in an exhaustive study mapping the human TNF-a/ NF-kB signal transduction network [76] , a role for the ER-to-Golgi vesicular pathway in RLH-induced innate immune response was still unknown until now. Future characterization of the TRAF3 interactome will undoubtedly help to understand the dA:dT (1 mg/ml) or SeV (200 HAU/ml) for 6 h to 8 h. RNA was extracted and analyzed by RT-qPCR using primers for ifnb, ifit1, oas1. Data are means +/ 2 S.D. (n = 3). * Significantly below the induction response; * P,0.05, ** P,0.01, *** P,0.001. (B and D) Cellular extracts were also prepared and subjected to immunoblot analysis using indicated antibodies. One of three independent experiments with similar results is shown. doi:10.1371/journal.ppat.1002747.g009 Figure 10 . p115 and Sec16A are required for optimal IRF-3 activation in response to activation of cytosolic RNA and DNA sensors. HeLa cells were infected with lentiviral vectors encoding shRNA targeting p115 (A) or Sec16A (B) and non-targeting (NT) control shRNA for 24 h followed by puromycin selection (1.5 mg/ml) for 5 days. Cells were left untreated or stimulated with poly I:C (1 mg/ml), poly dA:dT (1 mg/ml) or SeV (200 HAU/ml) for 16 h. Whole-cell lysates were prepared and subjected to immunoblot analysis with indicated antibodies. One of two independent experiments with similar results is shown. doi:10.1371/journal.ppat.1002747.g010 molecular relevance of the specific subcellular localization of TRAF3 for an optimal type I IFN response.