Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response.
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. Furthermo...
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Public Library of Science (PLoS)
2014
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oai:doaj.org-article:f96658edd07f4b1fb196fa2464f3ef812021-11-18T06:06:55ZHuman genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response.1553-73661553-737410.1371/journal.ppat.1003981https://doaj.org/article/f96658edd07f4b1fb196fa2464f3ef812014-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24586175/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374The 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.Niyas Kudukkil PulloorSajith NairKathleen McCaffreyAleksandar D KosticPradeep BistJeremy D WeaverAndrew M RileyRicha TyagiPradeep D UchilJohn D YorkSolomon H SnyderAdolfo García-SastreBarry V L PotterRongtuan LinStephen B ShearsRamnik J XavierManoj N KrishnanPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 10, Iss 2, p e1003981 (2014) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 |
| spellingShingle |
Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Niyas Kudukkil Pulloor Sajith Nair Kathleen McCaffrey Aleksandar D Kostic Pradeep Bist Jeremy D Weaver Andrew M Riley Richa Tyagi Pradeep D Uchil John D York Solomon H Snyder Adolfo García-Sastre Barry V L Potter Rongtuan Lin Stephen B Shears Ramnik J Xavier Manoj N Krishnan Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| description |
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. |
| format |
article |
| author |
Niyas Kudukkil Pulloor Sajith Nair Kathleen McCaffrey Aleksandar D Kostic Pradeep Bist Jeremy D Weaver Andrew M Riley Richa Tyagi Pradeep D Uchil John D York Solomon H Snyder Adolfo García-Sastre Barry V L Potter Rongtuan Lin Stephen B Shears Ramnik J Xavier Manoj N Krishnan |
| author_facet |
Niyas Kudukkil Pulloor Sajith Nair Kathleen McCaffrey Aleksandar D Kostic Pradeep Bist Jeremy D Weaver Andrew M Riley Richa Tyagi Pradeep D Uchil John D York Solomon H Snyder Adolfo García-Sastre Barry V L Potter Rongtuan Lin Stephen B Shears Ramnik J Xavier Manoj N Krishnan |
| author_sort |
Niyas Kudukkil Pulloor |
| title |
Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| title_short |
Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| title_full |
Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| title_fullStr |
Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| title_full_unstemmed |
Human genome-wide RNAi screen identifies an essential role for inositol pyrophosphates in Type-I interferon response. |
| title_sort |
human genome-wide rnai screen identifies an essential role for inositol pyrophosphates in type-i interferon response. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2014 |
| url |
https://doaj.org/article/f96658edd07f4b1fb196fa2464f3ef81 |
| work_keys_str_mv |
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