# Mechanisms and biological functions of SPOUT methyltransferases

> **NIH NIH R01** · OHIO STATE UNIVERSITY · 2024 · $48,062

## Abstract

PROJECT SUMMARY/ ABSTRACT
Transfer RNAs (tRNAs) are the universal adaptor molecules necessary to convert the nucleic acid-based genetic
code into protein sequence during protein synthesis (translation) by the ribosome. This process is universally
conserved and fundamental to all life, and, as such, defects in the molecular players of translation, including
tRNAs, result in diverse human diseases. Specific chemical modifications such as methylation are common in
tRNA, but a detailed understanding of the enzymes that incorporate them and their contributions to tRNA function
(and disfunction in disease) have only recently emerged for a few select examples. Since the discovery of the
tRNA methyltransferase (Trm10) in Saccharomyces cerevisiae, an accumulating body of evidence, including
phenotypes in yeast and a multisymptomatic disease associated with human mutations, has established a
significant role for Trm10 in tRNA biology. To better understand the implications of Trm10 modification, the
mechanisms by which Trm10 family enzymes specifically recognize and act on their substrate tRNA, and the
impact of tRNA modifications on important cellular processes need to be addressed. This project will determine
the molecular basis for Trm10 mechanism and function using a multi-disciplinary approach. Genetic, biochemical
and molecular enzymology approaches will be combined with structural analyses of enzyme-tRNA complexes
using synthetic analogs of the native methyl donor, S-adenosyl-L-methionine, to uniquely identify the role of
Trm10 in the maintenance of a high-quality pool of tRNA. A newly developed vertebrate model for Trm10 function
will enable investigation of previously challenging questions on Trm10's role in the biological function of
multicellular eukaryotes. The studies will be performed in three complementary but independent aims that will:
1) Determine how specific tRNA substrates are selected for modification by yeast and vertebrate Trm10 enzymes
using structural, biochemical and genetic approaches; 2) Assess the molecular basis for and biological
significance of the uniquely conserved vertebrate m1A9 modification exploiting a new vertebrate model for Trm10
function, and 3) Identify tRNA-specific functions for G9 modification in yeast and zebrafish using complementary
genetic approaches in both model species. Collectively, the proposed studies will advance the fields of
enzymology, RNA biochemistry, and tRNA biology by providing mechanistic and biological insight into a tRNA
modification enzyme that is universally conserved among eukaryotes and is critically important for human health,
yet whose molecular mechanism and biological functions are not at all understood. These results will also provide
new insight into the dynamic landscape of tRNA modifications in multicellular eukaryotes.

## Key facts

- **NIH application ID:** 11034339
- **Project number:** 3R01GM130135-05S1
- **Recipient organization:** OHIO STATE UNIVERSITY
- **Principal Investigator:** Graeme L Conn
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $48,062
- **Award type:** 3
- **Project period:** 2018-09-04 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11034339

## Citation

> US National Institutes of Health, RePORTER application 11034339, Mechanisms and biological functions of SPOUT methyltransferases (3R01GM130135-05S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/11034339. Licensed CC0.

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