Changes to the global transcriptome in human U2OS and BJ cell lines as a result of STING activation

Activation of Stimulator of interferon genes (STING) is well known to upregulate inflammation through canonical TBK1 signaling. STING is recently found to activate noncanonical functions through its transmembrane proton channel. Whether STING stimulates any transcription programs through its channel is unknown. In this study, we identified the transcription factor EB (TFEB) as a downstream effector of the STING channel. TFEB is a transcriptional regulator of lysosomal biogenesis and autophagy. Using RNA sequencing approaches, we confirmed that STING activated transcriptional upregulation of lysosomal biogenesis and autophagy independently of TBK1. Two sets of experiments were performed. First, U2OS cells stably expressing mouse STING1-350, which cannot activate TBK1, were treated with or without DMXAA for 8 hours, and the total RNAs were isolated for sequencing. Second, BJ WT and STING-KO cells were treated with digitonin buffer with or without 1 uM cGAMP, and the total RNAs were isolated for sequencing.