Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response.

Published

Journal Article

The 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.

Full Text

Duke Authors

Cited Authors

  • Pulloor, NK; Nair, S; McCaffrey, K; Kostic, AD; Bist, P; Weaver, JD; Riley, AM; Tyagi, R; Uchil, PD; York, JD; Snyder, SH; García-Sastre, A; Potter, BVL; Lin, R; Shears, SB; Xavier, RJ; Krishnan, MN

Published Date

  • February 27, 2014

Published In

Volume / Issue

  • 10 / 2

Start / End Page

  • e1003981 -

PubMed ID

  • 24586175

Pubmed Central ID

  • 24586175

Electronic International Standard Serial Number (EISSN)

  • 1553-7374

International Standard Serial Number (ISSN)

  • 1553-7366

Digital Object Identifier (DOI)

  • 10.1371/journal.ppat.1003981

Language

  • eng