tRNA-derived fragments (tRFs) regulate post-transcriptional gene expression via AGO-dependent mechanism in IL-1β stimulated chondrocytes.

Autor: Green JA; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH, 44272, USA., Ansari MY; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH, 44272, USA., Ball HC; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH, 44272, USA., Haqqi TM; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH, 44272, USA. Electronic address: thaqqi@neomed.edu.
Jazyk: angličtina
Zdroj: Osteoarthritis and cartilage [Osteoarthritis Cartilage] 2020 Aug; Vol. 28 (8), pp. 1102-1110. Date of Electronic Publication: 2020 May 12.
DOI: 10.1016/j.joca.2020.04.014
Abstrakt: 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-Cys GCA 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-Cys GCA . tRF-3003a "seed sequence" was identified in the 3'UTR of JAK3 mRNA and tRNA-Cys GCA 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.
(Copyright © 2020 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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