Abstract
Type II interferon (IFNγ) signaling is essential for innate immunity and critical for effective immunological checkpoint blockade in cancer immunotherapy. Genetic screen identification of post-transcriptional regulators of this pathway has been challenging since such factors are often essential for cell viability. Here, we utilize our inducible CRISPR/Cas9 approach to screen for key post-transcriptional regulators of IFNγ signaling, and in this way, we identify ERH and the ERH-associated splicing and RNA export factors MAGOH, SRSF1, and ALYREF. Loss of these factors impairs post-transcriptional mRNA maturation of JAK2, a crucial kinase for IFNγ signaling, resulting in abrogated JAK2 protein levels and diminished IFNγ signaling. Further analysis highlights a critical role for ERH in preventing intron retention in AU-rich regions in specific transcripts, such as JAK2. This regulation is markedly different from previously described retention of GC-rich introns. Overall, these findings reveal that post-transcriptional JAK2 processing is a critical rate-limiting step for the IFNγ-driven innate immune response.
| Originalsprache | Englisch |
|---|---|
| Aufsatznummer | gkaf545 |
| Fachzeitschrift | Nucleic Acids Research |
| Jahrgang | 53 |
| Ausgabenummer | 12 |
| DOIs | |
| Publikationsstatus | Veröffentlicht - 20 Juni 2025 |
Fördermittel
All RNA-seq was performed by the Next Generation Sequencing Facility at the Vienna BioCenter Core Facilities (VBCF), member of the Vienna BioCenter (VBC), Austria. Cell sorting for the genetic screens was performed at the IMP BioOptics flow cytometry facility. All other flow cytometry analyses were performed at the BioOptics FACS Facility at the Max Perutz Labs using the Max Perutz Labs instrument pool; we particularly acknowledge Kitti Csalyi, Thomas Sauer, and Johanna Stranner for expert support. Microscopy was performed at the Max Perutz Labs; we thank Irmgard Fischer for her expert support and training. We thank Michael P. Rout for providing a LaG16 nanobody plasmid, and the Protein Production Core Facility (PPCF), IMB, Mainz, Germany for GFP-trap beads preparation. We thank Niels Gehring, Volker B\u00F6hm, Sebastian Falk, and Clemens Plaschka for expert advice, discussions, and feedback on the manuscript. We are grateful to the Signaling Mechanisms in Cellular Homeostasis doctoral program community, in particular Thomas Decker, Pavel Kovarik, and their lab members. We are grateful to Laura Boccuni for technical expertise and help. We thank Life Science Editors for editing services. This research was funded in whole, or in part, by the Austrian Science Fund (FWF) (grants 10.55776/P36572, 10.55776/P30415, 10.55776/P30231, 10.55776/P36945, 10.55776/F79, and 10.55776/W1261 to G.A.V.). For the purpose of open access, the author has applied a CC-BY public copyright license to any author accepted manuscript version arising from this submission. M.V., M.deA., and S.S. are the recipients of a DOC fellowship of the Austrian Academy of Sciences. Research at the IMP is supported by Boehringer Ingelheim and the Austrian Research Promotion Agency (Headquarter grant FFG-852936). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. \u00D6sterreichische Forschungsf\u00F6rderungsgesellschaft, (Grant / Award Number: FFG-852936 ). Funding to pay the Open Access publication charges for this article was provided by Austrian Science Fund. This research was funded in whole, or in part, by the Austrian Science Fund (FWF) (grants 10.55776/P36572, 10.55776/P30415, 10.55776/P30231, 10.55776/P36945, 10.55776/F79, and 10.55776/W1261 to G.A.V.). For the purpose of open access, the author has applied a CC-BY public copyright license to any author accepted manuscript version arising from this submission. M.V., M.deA., and S.S. are the recipients of a DOC fellowship of the Austrian Academy of Sciences. Research at the IMP is supported by Boehringer Ingelheim and the Austrian Research Promotion Agency (Headquarter grant FFG-852936). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. \u00D6sterreichische Forschungsf\u00F6rderungsgesellschaft, (Grant / Award Number: \u2018FFG-852936\u2019). Funding to pay the Open Access publication charges for this article was provided by Austrian Science Fund.
ÖFOS 2012
- 106023 Molekularbiologie
UN SDGs
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