Poster abstracts
Poster number 101 submitted by Aftab Mollah
Unveiling the sequence-specific recognition of N6-methyladenosine (m6A) in eukaryotic RNAs
Aftab Mollah (Kent State University), Dr. Rushdhi Rauf (Kent State University), Dr. Sanjaya Abeysirigunawardena (Kent State University)
Abstract:
The prevalence of N6-methyladenosine (m6A) as the primary nucleotide modification in eukaryotic RNAs underscores its pivotal role in post-transcriptional regulation of gene expression. However, the intricate molecular mechanisms governing m6A recognition by various readers remain elusive. To address this gap, we conducted phage display experiments aimed at deciphering the essential sequence requirements for m6A binding. Strikingly, the same peptide (MP1) was selected for several RNA constructs harboring m6A, emphasizing the significance of specific protein sequence motifs in m6A recognition. Complementary RNA pulldown assays coupled with mass spectrometry revealed proteins with that bind specifically to methylated RNA that showed sequence similarity to peptides enriched in phase display experiments suggesting the importance of protein sequence in m6A recognition. These enriched proteins shared sequence and structural similarities with established m6A readers. Further validation through calorimetric methods highlight the importance of distinctive sequence features of proteins that govern the recognition of N6-methyl modifications. Our study contributes valuable insights into the molecular basis of m6A recognition, with potential implications for understanding broader regulatory networks in gene expression.
Keywords: RNA-Protein Interaction, m6A, Epitranscriptomic Regulation