In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress

Abstract Transfer RNA (tRNA) is the most abundant cellular RNA family in terms of copy numbers. It not only folds into defined structures but also has complex cellular interaction networks involving aminoacyl-tRNA synthetases, translation factors, and ribosomes. The human tRNAome is comprised of chr...

Full description

Saved in:
Bibliographic Details
Main Authors: Noah Peña, Yichen Hou, Christopher P. Watkins, Sihao Huang, Wen Zhang, Christopher D. Katanski, Tao Pan
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59435-5
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849704664954568704
author Noah Peña
Yichen Hou
Christopher P. Watkins
Sihao Huang
Wen Zhang
Christopher D. Katanski
Tao Pan
author_facet Noah Peña
Yichen Hou
Christopher P. Watkins
Sihao Huang
Wen Zhang
Christopher D. Katanski
Tao Pan
author_sort Noah Peña
collection DOAJ
description Abstract Transfer RNA (tRNA) is the most abundant cellular RNA family in terms of copy numbers. It not only folds into defined structures but also has complex cellular interaction networks involving aminoacyl-tRNA synthetases, translation factors, and ribosomes. The human tRNAome is comprised of chromosomal-encoded tRNAs with a large sequence diversity and mitochondrial-encoded tRNAs with A/U-rich sequences and noncanonical tertiary interactions. How tRNA folding and interactions in a eukaryotic cell respond to stress is poorly understood. Here, we develop DM-DMS-MaPseq, which utilizes in vivo dimethyl-sulfate (DMS) chemical probing and mutational profiling (MaP) coupled with demethylase (DM) treatment in transcriptome-wide tRNA sequencing to profile structures and the cellular interactions of human chromosomal and mitochondrial-encoded tRNAs. We found that tRNAs maintain stable structures in vivo, but the in vivo DMS profiles are vastly different from those in vitro, which can be explained by their interactions with cellular proteins and the ribosome. We also identify cytosolic and mitochondrial tRNA structure and interaction changes upon arsenite treatment, a type of oxidative stress that induces translational reprogramming, which is consistent with global translation repression in both compartments. Our results reveal variations of tRNA structurome and dynamic interactome that have functional consequences in translational regulation.
format Article
id doaj-art-00cfed91da224aa5972fea7b18bec037
institution DOAJ
issn 2041-1723
language English
publishDate 2025-05-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-00cfed91da224aa5972fea7b18bec0372025-08-20T03:16:41ZengNature PortfolioNature Communications2041-17232025-05-0116111610.1038/s41467-025-59435-5In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stressNoah Peña0Yichen Hou1Christopher P. Watkins2Sihao Huang3Wen Zhang4Christopher D. Katanski5Tao Pan6Department of Molecular Genetics and Cell Biology, University of ChicagoCommittee on Genomics, Genetics, and Systems Biology, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoCommittee on Genomics, Genetics, and Systems Biology, University of ChicagoAbstract Transfer RNA (tRNA) is the most abundant cellular RNA family in terms of copy numbers. It not only folds into defined structures but also has complex cellular interaction networks involving aminoacyl-tRNA synthetases, translation factors, and ribosomes. The human tRNAome is comprised of chromosomal-encoded tRNAs with a large sequence diversity and mitochondrial-encoded tRNAs with A/U-rich sequences and noncanonical tertiary interactions. How tRNA folding and interactions in a eukaryotic cell respond to stress is poorly understood. Here, we develop DM-DMS-MaPseq, which utilizes in vivo dimethyl-sulfate (DMS) chemical probing and mutational profiling (MaP) coupled with demethylase (DM) treatment in transcriptome-wide tRNA sequencing to profile structures and the cellular interactions of human chromosomal and mitochondrial-encoded tRNAs. We found that tRNAs maintain stable structures in vivo, but the in vivo DMS profiles are vastly different from those in vitro, which can be explained by their interactions with cellular proteins and the ribosome. We also identify cytosolic and mitochondrial tRNA structure and interaction changes upon arsenite treatment, a type of oxidative stress that induces translational reprogramming, which is consistent with global translation repression in both compartments. Our results reveal variations of tRNA structurome and dynamic interactome that have functional consequences in translational regulation.https://doi.org/10.1038/s41467-025-59435-5
spellingShingle Noah Peña
Yichen Hou
Christopher P. Watkins
Sihao Huang
Wen Zhang
Christopher D. Katanski
Tao Pan
In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
Nature Communications
title In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
title_full In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
title_fullStr In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
title_full_unstemmed In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
title_short In vivo structure profiling reveals human cytosolic and mitochondrial tRNA structurome and interactome in response to stress
title_sort in vivo structure profiling reveals human cytosolic and mitochondrial trna structurome and interactome in response to stress
url https://doi.org/10.1038/s41467-025-59435-5
work_keys_str_mv AT noahpena invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT yichenhou invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT christopherpwatkins invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT sihaohuang invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT wenzhang invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT christopherdkatanski invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress
AT taopan invivostructureprofilingrevealshumancytosolicandmitochondrialtrnastructuromeandinteractomeinresponsetostress