Isolation and initial structure-functional characterization of endogenous tRNA-derived stress-induced RNAs

• Published in: RNA biology Vol. 17; no. 8; pp. 1116 - 1124
• Main Authors: Akiyama, Yasutoshi, Kharel, Prakash, Abe, Takaaki, Anderson, Paul, Ivanov, Pavel
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 02.08.2020
• Subjects: angiogenin; ribonuclease; tRNA; tRNA fragments; tRNA-derived stress-induced RNAs; angiogenin; tRNA; tRNA fragments; tRNA-derived stress-induced RNAs; ribonuclease
• ISSN: 1547-6286; 1555-8584
• DOI: 10.1080/15476286.2020.1732702

Recent transcriptome-wide studies have identified a diverse pool of transfer RNA (tRNA)-derived RNAs or tRNA-derived fragments (tRFs). Some of these RNAs have been demonstrated to be functional and involved in multiple biological processes ranging from the regulation of gene expression to transgenerational epigenetic inheritance. Post-transcriptional maturation of tRNAs includes various processing events including extensive decoration by various RNA modifications, which are required for correct tRNA folding and stability. Moreover, tRNA modifications determine the pattern and specificity of tRNA cleavage. The major drawbacks of many studies in the field of tRFs are that most of them used synthetic RNAs that closely mimic endogenous tRFs in their sequence, yet lack RNA modification that is found in vivo. Here, we developed a simple method to isolate tRNA-derived stress-induced RNAs (tiRNAs), a specific subset of tRFs. Our approach is scalable, cost-effective and relies on the purification of individual tiRNAs based on a sequence-specific RNA/DNA isolation technique using DNA probes. Our method facilitates functional studies of tiRNAs by addressing how physiological RNA modifications within tRNA fragments affect their biological activities. Here, we report pilot functional studies on selected endogenous tiRNAs, namely tiRNA Ala and tiRNA Gly . We show that natural 5ʹ-tiRNA Ala molecules assemble into G-quadruplex structures, and endogenous 5ʹ-tiRNA Gly possesses translation inhibition activity.