Pulloor, Niyas KudukkilNair, SajithKostic, AleksandarBist, PradeepWeaver, Jeremy D.Riley, Andrew M.Tyagi, RichaUchil, Pradeep D.York, John D.Snyder, Solomon H.García-Sastre, AdolfoPotter, Barry V. L.Lin, RongtuanShears, Stephen B.Xavier, RamnikKrishnan, Manoj N.2014-04-112014Pulloor, N. K., S. Nair, A. D. Kostic, P. Bist, J. D. Weaver, A. M. Riley, R. Tyagi, et al. 2014. “Human Genome-Wide RNAi Screen Identifies an Essential Role for Inositol Pyrophosphates in Type-I Interferon Response.” PLoS Pathogens 10 (2): e1003981. doi:10.1371/journal.ppat.1003981. http://dx.doi.org/10.1371/journal.ppat.1003981.1553-7366http://nrs.harvard.edu/urn-3:HUL.InstRepos:12064389The pattern recognition receptor RIG-I is critical for Type-I interferon production. However, the global regulation of RIG-I signaling is only partially understood. Using a human genome-wide RNAi-screen, we identified 226 novel regulatory proteins of RIG-I mediated interferon-β production. Furthermore, the screen identified a metabolic pathway that synthesizes the inositol pyrophosphate 1-IP7 as a previously unrecognized positive regulator of interferon production. Detailed genetic and biochemical experiments demonstrated that the kinase activities of IPPK, PPIP5K1 and PPIP5K2 (which convert IP5 to1-IP7) were critical for both interferon induction, and the control of cellular infection by Sendai and influenza A viruses. Conversely, ectopically expressed inositol pyrophosphate-hydrolases DIPPs attenuated interferon transcription. Mechanistic experiments in intact cells revealed that the expression of IPPK, PPIP5K1 and PPIP5K2 was needed for the phosphorylation and activation of IRF3, a transcription factor for interferon. The addition of purified individual inositol pyrophosphates to a cell free reconstituted RIG-I signaling assay further identified 1-IP7 as an essential component required for IRF3 activation. The inositol pyrophosphate may act by β-phosphoryl transfer, since its action was not recapitulated by a synthetic phosphonoacetate analogue of 1-IP7. This study thus identified several novel regulators of RIG-I, and a new role for inositol pyrophosphates in augmenting innate immune responses to viral infection that may have therapeutic applications.en-USBiologyImmunologyImmunityInnate immunityImmune responseMedicineInfectious diseasesViral diseasesJournal Article2014-04-1110.1371/journal.ppat.1003981