Poster abstracts
Poster number 72 submitted by David Morgan
Identifying novel m6A binding proteins using a peptide as a bait which was enriched in a phage display experiment.
David Morgan (Department of Chemistry and Biochemistry - Kent State University ), Rushdhi Rauff (Department of Chemistry and Biochemistry - Kent State University ), Sanjaya Abeysirigunawardena (Department of Chemistry and Biochemistry - Kent State University )
Abstract:
N6-methyladenosine (m6A) is the most abundant nucleotide modification in eukaryotic mRNA. The m6A modification can be added, removed, and recognized by m6A writers, erasers, and readers, respectively. Due to the presence of many m6A modifications in some mRNAs, likely there are more m6A binding proteins that can interact site-specifically. Protein pull-down assays were performed using various methylated and unmethylated RNA targets to identify proteins that are bound to methylated RNA with sequence and structural specificity. Lysates from various cancer cell-lines were used for these pull-down assays. Interestingly, the enriched proteins have sequence similarity to dodecamer peptides selected from phage display experiments using same RNA baits. Our experiments show that m6A readers interacts with RNA using unique strategies.
Keywords: m6A, Nucleotide modifications, Epitranscriptomics