Poster abstracts
Poster number 47 submitted by Chathurani Ekanayake
Investigation of the ability of 5-methylcytosine to modulate RNA-protein interactions in humans
Chathurani Ekanayake (Department of Chemistry and Biochemistry, Kent State University, Kent OH), Sanjaya Abeysirigunawardena (Department of Chemistry and Biochemistry, Kent State University, Kent OH)
Abstract:
During the epitranscriptomic regulation of gene expression, post-transcriptional modifications such as m5C, m7G, m1A, and Ψ play crucial roles in functional RNAs. Dysregulation of mRNA nucleotide modifications is associated with various human diseases, including cancer, diabetes, and neurodevelopmental disorders. Despite their prevalence, the specific mechanisms underlying RNA modification recognition and their role in disease pathogenesis remain incompletely understood. This study aims to investigate the impact of RNA sequence on m5C recognition by proteins. Phage display experiments were conducted using several m5C containing model RNAs with diverse sequences, including consensus sequences for NSUN2 and NSUN6 methyltransferases. The impact of RNA sequence on m5C recognition by proteins was examined, particularly in the breast cancer cell line MDA-MB 231. RNA pull-down assays targeting NSUN6, NSUN6 M5C, NSUN2, and NSUN2 M5C were performed, followed by LC-MS/MS analysis to identify associated proteins. Peptide alignment analysis revealed specific interactions between peptides and protein structures, suggesting a potential methyl-reading function. Results indicated enrichment of RNA-binding proteins, highlighting a complex regulatory network involved in m5C recognition. Protein Network analyses provided insights into functional implications and pathways influenced by the identified proteins. These findings enhance our understanding of RNA modification recognition mechanisms and their dysregulation in breast cancer. Future research will focus on the functional characterization of identified proteins and their implications in disease pathogenesis, potentially leading to targeted therapeutic interventions. Overall, this study contributes to elucidating the molecular mechanisms underlying gene expression regulation and offers insights into potential therapeutic targets in breast cancer.
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Keywords: RNA-protein interactions, m5C methylation, Breast cancer