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Gene Synthesis> | pRK5 plasmids encoding hemagglutinin (HA)–tagged ubiquitin wild-type (wt), K48, and K63 were provided by T. Dawson (Addgene plasmids #17605, #17606, and #17608) (51). Flag-tagged IRF7 wt was purchased from GenScript | Get A Quote |
Plasmacytoid dendritic cells (pDCs) play a crucial role in antiviral innate immunity through their unique capacity to produce large amounts of type I interferons (IFNs) upon viral detection. Tripartite motif (TRIM) proteins have recently come forth as important modulators of innate signaling, but their involvement in pDCs has not been investigated. Here, we performed a rationally streamlined small interfering RNA (siRNA)-based screen of TRIM proteins in human primary pDCs to identify those that are critical for the IFN response. Among candidate hits, TRIM8 emerged as an essential regulator of IFN regulatory factor 7 (IRF7) function. Mechanistically, TRIM8 protects phosphorylated IRF7 (pIRF7) from proteasomal de... More