tRNA-derived fragments (tRFs) regulate post-transcriptional gene expression via AGO-dependent mechanism in IL-1β stimulated chondrocytes
| Autor: | Jonathan A Green, Tariq M. Haqqi, Hope C. Ball, Mohammad Y. Ansari |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Interleukin-1beta Primary Cell Culture Biomedical Engineering Article Chondrocyte Cell Line 03 medical and health sciences Chondrocytes 0302 clinical medicine RNA Transfer Rheumatology Osteoarthritis Gene expression medicine Humans Gene silencing Orthopedics and Sports Medicine RNA Messenger 3' Untranslated Regions 030203 arthritis & rheumatology Regulation of gene expression RNA Transfer Cys Gene knockdown Messenger RNA Chemistry Janus Kinase 3 RNA Transfection Cell biology 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation Argonaute Proteins RNA Small Untranslated |
| Zdroj: | Osteoarthritis Cartilage |
| ISSN: | 1063-4584 |
| DOI: | 10.1016/j.joca.2020.04.014 |
| Popis: | Summary Objectives Recent studies have shown that tRNA-derived RNA fragments (tRFs) are novel regulators of post-transcriptional gene expression. However, the expression profiles and their role in post-transcriptional gene regulation in chondrocytes is unknown. Here, we determined tRFs expression profile and explored tRF-3003a role in post-transcriptional gene regulation in IL-1β stimulated chondrocytes. Methods We used qPCR arrays to determine tRNAs and tRFs expression in age- and sex-matched primary human OA chondrocytes and TC28/I2 cells stimulated with IL-1β. Chondrocytes were transfected with tRNA-CysGCA overexpression plasmid or tRF-3003a mimic and 3′UTR luciferase reporter plasmids of mRNAs harboring predicted tRF target “seed sequence”. The AGO-RNA-induced silencing complex (AGO-RISC)-dependent repressive activity of tRF-3003a was determined by siRNA-mediated knockdown of AGO2. Results IL-1β increased the expression levels of specific tRNAs and of tRF-3003a, a type 3 tRF produced by the cleavage of tRNA-CysGCA. tRF-3003a “seed sequence” was identified in the 3′UTR of JAK3 mRNA and tRNA-CysGCA overexpression or transfection of a tRF-3003a mimic in chondrocytes downregulated JAK3 expression and significantly reduced the activity of the 3′UTR reporter. RIP assay showed enrichment of tRF-3003a into AGO2/RISC in IL-1β treated chondrocytes. The suppressive effect of tRF-3003a on JAK3 3′UTR reporter was abrogated with siRNA-mediated depletion of AGO2. Conclusions We demonstrate that under pathological conditions chondrocytes display perturbations in the expression profile of specific tRNAs and tRFs. Furthermore, a specific tRF namely tRF-3003a can post-transcriptionally regulate JAK3 expression via AGO/RISC formation in chondrocytes. Identification of this novel mechanism may be of value in the design of precision therapies for OA. |
| Databáze: | OpenAIRE |
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