A multiplex platform for small RNA sequencing elucidates multifaceted tRNA stress response and translational regulation

Christopher P. Watkins, Wen Zhang, Adam C. Wylder, Christopher D. Katanski () and Tao Pan ()
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Christopher P. Watkins: Department of Biochemistry and Molecular Biology
Wen Zhang: University of Chicago
Adam C. Wylder: University of Chicago
Christopher D. Katanski: Department of Biochemistry and Molecular Biology
Tao Pan: Department of Biochemistry and Molecular Biology

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Small RNAs include tRNA, snRNA, micro-RNA, tRNA fragments and others that constitute > 90% of RNA copy numbers in a human cell and perform many essential functions. Popular small RNA-seq strategies limit the insights into coordinated small RNA response to cellular stress. Small RNA-seq also lacks multiplexing capabilities. Here, we report a multiplex small RNA-seq library preparation method (MSR-seq) to investigate cellular small RNA and mRNA response to heat shock, hydrogen peroxide, and arsenite stress. Comparing stress-induced changes of total cellular RNA and polysome-associated RNA, we identify a coordinated tRNA response that involves polysome-specific tRNA abundance and synergistic N3-methylcytosine (m3C) tRNA modification. Combining tRNA and mRNA response to stress we reveal a mechanism of stress-induced down-regulation in translational elongation. We also find that native tRNA molecules lacking several modifications are biased reservoirs for the biogenesis of tRNA fragments. Our results demonstrate the importance of simultaneous investigation of small RNAs and their modifications in response to varying biological conditions.

Date: 2022
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DOI: 10.1038/s41467-022-30261-3

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