RTCB Complex Regulates Stress-Induced tRNA Cleavage
Abstract
:1. Introduction
2. Results
2.1. RTCB Ligase Complex Negatively Regulates tiRNA Production
2.2. tiRNAs Can Be Repaired to Full Length tRNAs In Vitro
2.3. Chimeric tRNAs Are Not Detected in the Cell
2.4. tiRNA Production Is Enhanced under Oxidative Stress Conditions through RTCB Inhibition
2.5. Inhibition of RTCB by Oxidation Is a Major Regulator of tiRNA Production
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Knockdown of RTCB
4.3. Western Blotting
4.4. In lysate Angiogenin Digestion
4.5. Northern Blotting
4.6. Ligation of tiRNAs by Bacterial RtcB Ligase
4.7. Pulldown of 6xHis-Tagged Angiogenin
4.8. Small RNA-Seq Data Analysis for Chimeric tRNA Generation
4.9. Quantitative PCR
4.10. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Akiyama, Y.; Takenaka, Y.; Kasahara, T.; Abe, T.; Tomioka, Y.; Ivanov, P. RTCB Complex Regulates Stress-Induced tRNA Cleavage. Int. J. Mol. Sci. 2022, 23, 13100. https://doi.org/10.3390/ijms232113100
Akiyama Y, Takenaka Y, Kasahara T, Abe T, Tomioka Y, Ivanov P. RTCB Complex Regulates Stress-Induced tRNA Cleavage. International Journal of Molecular Sciences. 2022; 23(21):13100. https://doi.org/10.3390/ijms232113100
Chicago/Turabian StyleAkiyama, Yasutoshi, Yoshika Takenaka, Tomoko Kasahara, Takaaki Abe, Yoshihisa Tomioka, and Pavel Ivanov. 2022. "RTCB Complex Regulates Stress-Induced tRNA Cleavage" International Journal of Molecular Sciences 23, no. 21: 13100. https://doi.org/10.3390/ijms232113100