# Investigation of the key structural and sequence features essential for m6A recognition during post-transcriptional regulation of gene expression

> **NIH NIH R15** · KENT STATE UNIVERSITY · 2020 · $454,624

## Abstract

PROJECT SUMMARY
The existence of RNA nucleotide modifications in functional RNAs is known for many decades.
Several recent studies illustrate the transcriptome-wide presence of nucleotide modifications such
as pseudouridines, N6-methyladenosines (m6A), and 5-methylcytosines. The levels of nucleotide
modifications in mRNA are in tight equilibrium unless cells are under various stress conditions.
Changes in m6A levels in mRNA have shown to impact viral infections, sperm maturation, and
cancer progression. In cells, m6A levels are controlled by methyl writers and readers. These
proteins codes the stress signal on to mRNA transcripts, both post-, and co-transcriptionally.
Methyl readers that recognize methylations play a critical role in decoding stress signals and direct
mRNA to either getting edited, processed, degraded, or translated. Given the broader diversity of
mRNA methylation states under various stress conditions and in human diseases, an assemblage
of methyl readers that can read each unique stress signal should exist. The lack of general
structural and sequence consensus for methyl-recognizing proteins (reader or erasers) impedes
the discovery of novel regulation mechanisms by readers and erasers not known up to date. The
three short term goals of this project are 1) to discover sequence or structural consensus for short
peptides that interact with m6A, 2) to understand how RNA structure and sequence can change
the sequence and the structure of m6A-recognizing peptides, 3) to investigate the ability of
enriched peptides to inhibit reader and eraser protein.
In this research, the phage display method is used to discover a sequence or structural consensus
essential for m6A recognition. The impact of RNA structure and sequence on the sequence or
structure consensus will also be tested. Sequence comparison will be made between the peptides
enriched against methylated RNA targets (phage display) and proteins identified from pulldown
assays. Preliminary result of this research illustrates that 1) RNA methylations enhance
sequence-specific interactions of RNA with proteins, 2) two tryptophan residues that reside four
amino acid residues apart in the peptide may play a role in m6A recognition, and 3) RNA binding
sites of writer or eraser proteins have similar sequences as the selected peptides against
unmodified and modified RNA targets, respectively. Long term objectives of this project include
engineering unique designer proteins in which m6A-recognizing peptides (that binding sequence
specifically or structure specifically) are fused with proteins related to RNA processing,
localization, and degradations to use in treating human diseases.

## Key facts

- **NIH application ID:** 10114391
- **Project number:** 1R15GM137291-01A1
- **Recipient organization:** KENT STATE UNIVERSITY
- **Principal Investigator:** Sanjaya Chinthaka Abeysirigunawardena
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $454,624
- **Award type:** 1
- **Project period:** 2020-09-15 → 2023-09-14

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10114391

## Citation

> US National Institutes of Health, RePORTER application 10114391, Investigation of the key structural and sequence features essential for m6A recognition during post-transcriptional regulation of gene expression (1R15GM137291-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10114391. Licensed CC0.

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