# Mechanisms of Regulation of the m6A mRNA Methylation Machinery

> **NIH NIH R35** · YALE UNIVERSITY · 2022 · $418,750

## Abstract

Project Summary
 RNA modifications (the epitranscriptome) represent a mechanism of post-transcriptional gene expression
regulation and an emergent and exciting area of biology. N6-methyladenosine (m6A) is the most abundant post-
transcriptional modification in mRNA, detected across the eukaryotic kingdom, from yeast to humans. m6A has been
implicated in multiple molecular processes such as RNA splicing, RNA stability and miRNA processing and cellular
functions such as meiosis, cell proliferation and embryonic stem cell differentiation, as well as disease states. The
methyltransferases responsible for the m6A form a heterodimer in which METTL3 provides the catalytic activity. We
and others have found that METTL3 has a positive impact in proliferation, differentiation and cell survival. Thus, we
postulate that METTL3 represents a regulatory hub controlled by extra- and intracellular signals that allows
the cells to respond to developmental cues and specific metabolic contexts by modulating RNA metabolism.
Despite the rapid increase in our knowledge about the functions of m6A, there are many fundamental aspects of this
process that remain unknown. For example, it is yet to be determined if the activity of the methyltransferases is
regulated by upstream signaling pathways, and if this is the case, what are the mechanisms involved, and what are
the molecular and cellular consequences of this regulation in homeostasis and development? Another key issue is to
understand how the specificity of the methylation reaction is determined. We know that the recognition sequence for
m6A methylation consists of a very short sequence motif – but despite the abundance of this sequence in the
transcriptome only a small fraction of such motifs gets methylated. Additional RNA structural motifs or sequence
requirements have not been identified. To answer these broad questions, our lab is undertaking a multidisciplinary
approach that includes complementary projects in the areas of molecular and cell biology, biochemistry as well as
structural studies complemented with novel engineered mouse models. The program described here will allow us to
solve the mechanisms involved in the regulation of the m6A methylation pathway upon extracellular stimulation,
metabolic stress and development. Using state-of-the-art proteomic techniques, we will identify context-specific post-
translational modifications acquired by METTL3, and their impact on the activity and specificity of the enzyme. We will
complement these studies with structural techniques including X-ray crystallography and cryo-electron microscopy to
define the molecular consequences of regulatory events on protein complex formation and substrate recognition. In
parallel, we will use our recently developed mouse model that allows the inducible and tissue specific inactivation of
METTL3 to distinguish between catalytic and non-catalytic functions of METTL3 and to understand the role of the m6A
mark in stem cell self-renewal...

## Key facts

- **NIH application ID:** 10439740
- **Project number:** 5R35GM138185-03
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Claudio R Alarcon
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $418,750
- **Award type:** 5
- **Project period:** 2020-08-14 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10439740

## Citation

> US National Institutes of Health, RePORTER application 10439740, Mechanisms of Regulation of the m6A mRNA Methylation Machinery (5R35GM138185-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10439740. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*