Characterization of a novel nucleotide based second messenger system regulating innate immune responses
Virology
Final Report Abstract
Nucleic acids play a central role in the antiviral innate immune response. Viral nucleic acids can be sensed by pattern recognition receptors, which stimulate antiviral immunity though eliciting the expression of type-I interferons or other cytokines that are important to induce an antiviral state in adjacent cells or in a given tissue. Moreover, shorter nucleic acids are important signalling molecules that distribute signals or lead to activation of intracellular proteins. A prominent example for this function is the activity of cGAMP, a cyclic di-nucleotide that is syntehsised by cGAS and activates the cellular protein STING to initiate expression of type-I interferon and cytokines. Another well-characterized nucleotide that plays an important role in antiviral immunity is 2’5’OA, which is synthesised by OAS proteins upon sensing of viruses and activates RNAseL. We proposed to study the activity of di-adenosine-tetraphosphate (Ap4A) which is synthesised upon cell stress and in cells with mutations in tRNA aminoacyltransferases (particularly KRAS) or in the absence of Nudix hytrolase 2 (NUDT2), which degrades AP4A. To understand the activity of AP4A we systematically investigated macrophage responses to AP4A and other extracellularly added nucleotides and found that they induce a prominent cytokine response. In order to find receptors that were potentially involved in this response we focused on the P2Y receptor (P2YR) family. Time-resolved transcriptomics analysis showed that adenine and uridine-based nucleotides induce a specific, immediate and transient cytokine response through the MAPK signaling pathway regulation of AP-1 transcriptional activation. Using receptor trans-complementation, we identified a subset of P2Ys (P2Y1, P2Y2, P2Y6 and P2Y11) that governs the cytokine induction, while others (P2Y4, P2Y11, P2Y12, P2Y13 and P2Y14) directly induced antiviral responses. Notably, P2Y11 combined both activities and depletion or inhibition of this receptor in macrophages impaired both inflammatory and antiviral responses. Our study provides in-depth characterization of the underappreciated functions of P2Y receptors in innate immune processes. We propose that nucleotide signalling is an important aspect that may shape the immune response in tissue and be relevant for induction of immune responses and viral clearance.
Publications
-
Cross-species analysis of viral nucleic acid interacting proteins identifies TAOKs as innate immune regulators. Nature Communications, 12(1).
Pennemann, Friederike L.; Mussabekova, Assel; Urban, Christian; Stukalov, Alexey; Andersen, Line Lykke; Grass, Vincent; Lavacca, Teresa Maria; Holze, Cathleen; Oubraham, Lila; Benamrouche, Yasmine; Girardi, Enrico; Boulos, Rasha E.; Hartmann, Rune; Superti-Furga, Giulio; Habjan, Matthias; Imler, Jean-Luc; Meignin, Carine & Pichlmair, Andreas
-
Genotoxic stress in constitutive trisomies induces autophagy and the innate immune response via the cGAS-STING pathway. Communications Biology, 4(1).
Krivega, Maria; Stiefel, Clara M.; Karbassi, Sahar; Andersen, Line L.; Chunduri, Narendra K.; Donnelly, Neysan; Pichlmair, Andreas & Storchová, Zuzana
-
Multilevel proteomics reveals host perturbations by SARS-CoV-2 and SARS-CoV. Nature, 594(7862), 246-252.
Stukalov, Alexey; Girault, Virginie; Grass, Vincent; Karayel, Ozge; Bergant, Valter; Urban, Christian; Haas, Darya A.; Huang, Yiqi; Oubraham, Lila; Wang, Anqi; Hamad, M. Sabri; Piras, Antonio; Hansen, Fynn M.; Tanzer, Maria C.; Paron, Igor; Zinzula, Luca; Engleitner, Thomas; Reinecke, Maria; Lavacca, Teresa M. ... & Pichlmair, Andreas
-
NUDT2 initiates viral RNA degradation by removal of 5′-phosphates. Nature Communications, 12(1).
Laudenbach, Beatrice T.; Krey, Karsten; Emslander, Quirin; Andersen, Line Lykke; Reim, Alexander; Scaturro, Pietro; Mundigl, Sarah; Dächert, Christopher; Manske, Katrin; Moser, Markus; Ludwig, Janos; Wohlleber, Dirk; Kröger, Andrea; Binder, Marco & Pichlmair, Andreas
-
Two cGAS-like receptors induce antiviral immunity in Drosophila. Nature, 597(7874), 114-118.
Holleufer, Andreas; Winther, Kasper Grønbjerg; Gad, Hans Henrik; Ai, Xianlong; Chen, Yuqiang; Li, Lihua; Wei, Ziming; Deng, Huimin; Liu, Jiyong; Frederiksen, Ninna Ahlmann; Simonsen, Bine; Andersen, Line Lykke; Kleigrewe, Karin; Dalskov, Louise; Pichlmair, Andreas; Cai, Hua; Imler, Jean-Luc & Hartmann, Rune
-
Attenuation of SARS‐CoV‐2 replication and associated inflammation by concomitant targeting of viral and host cap 2'‐O‐ribose methyltransferases. The EMBO Journal, 41(17).
Bergant, Valter; Yamada, Shintaro; Grass, Vincent; Tsukamoto, Yuta; Lavacca, Teresa; Krey, Karsten; Mühlhofer, Maria‐Teresa; Wittmann, Sabine; Ensser, Armin; Herrmann, Alexandra; vom Hemdt, Anja; Tomita, Yuriko; Matsuyama, Shutoku; Hirokawa, Takatsugu; Huang, Yiqi; Piras, Antonio; Jakwerth, Constanze A.; Oelsner, Madlen; Thieme, Susanne ... & Kato, Hiroki
-
Systematic P2Y receptor survey identifies P2Y11 as modulator of immune responses and virus replication in macrophages. The EMBO Journal, 42(23).
Andersen, Line Lykke; Huang, Yiqi; Urban, Christian; Oubraham, Lila; Winheim, Elena; Stafford, Che; Nagl, Dennis; O.'Duill, Fionan; Ebert, Thomas; Engleitner, Thomas; Paludan, Søren Riis; Krug, Anne; Rad, Roland; Hornung, Veit & Pichlmair, Andreas
