2023 Fiscal Year Research-status Report
Structural and computational analysis of immune-related RNA-binding proteins
| Project/Area Number |
23K06597
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| Research Institution | Osaka University |
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Principal Investigator |
MILLIUS ARTHUR 大阪大学, 微生物病研究所, 特任研究員(常勤) (80624858)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Keywords | Ribosome profiling / Circadian rhythms / Period2 / Translation / uORF / RNA structure / Regnase / Machine learning |
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| Outline of Annual Research Achievements |
We are exploring how different RNA structures in 5' and 3' UTRs affect translation and RNA degradation in cells and animals to understand how post-transcriptional and translational mechanisms influence when and how much of a protein gets made. First, we used ribosome profiling to examine how upstream open reading frames control Period2 RNA processing in sleep and circadian rhythms (Millius et al., 2023), and now we have started to examine how an endoribonuclease Regnase-1 degrades RNAs in the immune system. Defects in circadian rhythms are related to health problems and T cell-specific targeting of Regnase-1 can be used as a cancer therapeutic, so a broad understanding of the cellular mechanisms that control RNA processing is essential for developing new treatments.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
In collaboration with the Ueda lab, we mutated the Per2 uORF using Crispr/Cas9 genetic editing and found that both male and female mutant mice had significantly reduced sleep compared to their wild-type littermates (Millius et al., 2023). We analyzed the 3’UTR of Regnase-1 to understand how Regnase-1 autoregulation controls its own expression (Piboonprai et al., 2023). Now we are screening chemical libraries to identify chemical inhibitors of Regnase-1.
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| Strategy for Future Research Activity |
We plan to crystallize Regnase wild-type and select Regnase mutant proteins in collaboration with Genji Kurisu's lab. We have generated new Regnase mouse models that affect the multimerization state of the Regnase protein, and we will examine how these mutations affect the physiology and immune system function of the mice. In addition, we are screening chemical libraries for potential inhibitors of Regnase binding to RNA.
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| Causes of Carryover |
In 2023, we focused on addressing comments from reviewers and publishing our results in Millius et al., 2023 and Piboonprai et al., 2023. Therefore, we had less time than anticipated for planning and implementation of our experiments. This year we plan to carry out the bulk of our studies on the structure of Regnase as well as screening chemical libraries for inhibitors of Regnase binding to RNA.
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