The article identifies and characterizes STING (STimulator of INterferon Genes), a molecule that regulates innate immune signaling processes. STING, composed of 5 putative transmembrane regions, primarily resides in the endoplasmic reticulum (ER) and activates both NF-κB and IRF3 transcription pathways to induce type I IFN, thereby exerting a potent antiviral state. Loss of STING in murine embryonic fibroblasts (MEFs) renders them highly susceptible to negative-stranded virus infection, including vesicular stomatitis virus (VSV). STING ablation also abrogates the ability of intracellular B-form DNA and herpes viruses to induce IFNβ but does not affect the Toll-like receptor (TLR) pathway. Yeast-two hybrid and co-immunoprecipitation studies indicate that STING interacts with RIG-I and Ssr2/TRAPβ, a member of the translocon-associated protein complex required for protein translocation across the ER membrane. RNAi ablation of TRAPβ and translocon adaptor Sec61β inhibits STING's ability to stimulate IFNβ, suggesting a role for the translocon in innate signaling pathways activated by select viruses and intracellular DNA. The study provides insights into the molecular mechanisms of STING's function in innate immune signaling and its potential role in linking RIG-I and DNA-mediated intracellular signaling to the translocon.The article identifies and characterizes STING (STimulator of INterferon Genes), a molecule that regulates innate immune signaling processes. STING, composed of 5 putative transmembrane regions, primarily resides in the endoplasmic reticulum (ER) and activates both NF-κB and IRF3 transcription pathways to induce type I IFN, thereby exerting a potent antiviral state. Loss of STING in murine embryonic fibroblasts (MEFs) renders them highly susceptible to negative-stranded virus infection, including vesicular stomatitis virus (VSV). STING ablation also abrogates the ability of intracellular B-form DNA and herpes viruses to induce IFNβ but does not affect the Toll-like receptor (TLR) pathway. Yeast-two hybrid and co-immunoprecipitation studies indicate that STING interacts with RIG-I and Ssr2/TRAPβ, a member of the translocon-associated protein complex required for protein translocation across the ER membrane. RNAi ablation of TRAPβ and translocon adaptor Sec61β inhibits STING's ability to stimulate IFNβ, suggesting a role for the translocon in innate signaling pathways activated by select viruses and intracellular DNA. The study provides insights into the molecular mechanisms of STING's function in innate immune signaling and its potential role in linking RIG-I and DNA-mediated intracellular signaling to the translocon.