# Structural and functional basis for protein-based eukaryotic RNA processing

> **NIH NIH R01** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2024 · $308,069

## Abstract

PROJECT ABSTRACT
Ribonucleic acids (RNAs) are amongst the most functionally versatile biomolecules found in nature,
accomplishing diverse biological tasks including carrying genetic information (mRNA, viral genomes),
catalyzing reactions (rRNA, RNase P, telomerase, group I/II introns), acting as molecular adaptors
(tRNA), and regulating gene expression (riboswitches, lncRNA, miRNA). Alterations in the structure and
dynamics of RNA molecules are linked to bacterial pathogenesis, cardiovascular conditions, neurological
disorders, mitochondrial diseases, and cancers. Despite the biological importance of RNA, our
understanding of fundamental aspects of RNA structure, function and biochemical properties lags far
behind that of proteins. My research program studies how the structure and dynamics of mitochondrial
transfer RNAs (mt-tRNAs) impact their maturation and function. Our work will fill long-standing
knowledge gaps in RNA biology–including identifying molecular level consequences of disease causing
mt-tRNA mutations (Aim 3), establishing the impact of discrete tRNA post-transcriptional modifications
(Aims 1&2), and determining the high-resolution structures of a broad set of RNAs and the proteins
involved in their maturation (Aims 1-3). Additionally, we will develop broadly applicable chemical tools
for studying mitochondrial RNA biology (Aim 3). Our preliminary work suggests that mt-tRNAs have
unique structural and biophysical properties, are often more dynamic and unstable prior to processing and
modification, and, therefore, have unique maturation pathways. We posit that disease-causing mutations
destabilize mt-tRNA structure and alter its ability to be recognized as a substrate by processing and
modification enzymes, therefore affecting mt-tRNA biogenesis and therefore mitochondrial function. In
this work we propose a combination of biochemistry (kinetic and binding assays), structural biology
(Cryo-EM and X-ray crystallography), innovative bio-analytical characterization (IM-MS), biophysical
tools (smFRET, NMR), chemical biology (high-throughput assays) and cellular biology to take on
questions regarding how mt-RNA structure and mt-PRORP function contribute to mitochondrial health
and pathology. These studies will reveal how tRNAs are recognized and processed by mt-PRORP and
help rationalize how patient-derived mt-tRNA mutations alter mt-tRNA stability and processing,
improving our understanding of fundamental aspects of mt-tRNA molecular biology and opening new
avenues of exploration into the role of mitochondrial post-transcriptional RNA modification in
mitochondrial diseases.

## Key facts

- **NIH application ID:** 10802717
- **Project number:** 2R01GM117141-07
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Markos Koutmos
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $308,069
- **Award type:** 2
- **Project period:** 2016-02-01 → 2028-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10802717

## Citation

> US National Institutes of Health, RePORTER application 10802717, Structural and functional basis for protein-based eukaryotic RNA processing (2R01GM117141-07). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10802717. Licensed CC0.

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