Poster abstracts
Poster number 37 submitted by Chathurani Ekanayake
Contrasting recognition mechanisms exists for m⁵C located withing different RNA sequences
Chathurani Ekanayake (Department of Chemistry and Biochemistry, Kent State University. ), Maelee Thompson (Department of Chemistry and Biochemistry, Kent State University. ), Aftab U. Mollah (Department of Chemistry and Biochemistry, Kent State University. ), Sanjaya C. Abeysirigunawardena (Department of Chemistry and Biochemistry, Kent State University.)
Abstract:
5-Methylcytosine (m⁵C) nucleotide modifications, which are abundant in eukaryotic mRNA, play a crucial role in regulating gene expression at both transcriptional and translational levels. Dysregulation of m⁵C has been linked to various diseases, including cancer, diabetes, and neurodevelopmental disorders. However, the mechanisms by which m⁵C modification is recognized by proteins are not fully understood. To address this gap, we performed phage display, a directed evolution method, to select peptides that specifically recognize m⁵C modifications in different sequence contexts. Two model RNAs with m⁵C modifications within sequences recognized by two methyltransferases, NSUN2 (ss-MN2) and NSUN6 (ss-MN6), were used as targets in these phage display experiments. Two distinct peptide pools were enriched against ss-MN2 and ss-MN6 RNAs, indicating contrasting recognition mechanisms for m⁵C located in different RNA sequences. Isothermal titration calorimetry (ITC) measurements revealed differing binding affinities for each enriched peptide, consistent with their percentage enrichment. Moving forward, a functional characterization of the identified proteins and their binding properties to various RNAs will be conducted to elucidate their recognition mechanisms. These insights could highlight the potential of targeting the m⁵C modification mechanism as a novel approach for drug development against various diseases.
Keywords: neucleotide modifications, RNA, Protein