An Investigation of the Interplay Between N6-Methyleadenosine (m6A), microRNAs and RNA Binding Proteins in Regulating mRNA Function

Saren, Soumyendu Narayan (2021) An Investigation of the Interplay Between N6-Methyleadenosine (m6A), microRNAs and RNA Binding Proteins in Regulating mRNA Function. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Soumyendu Narayan Saren (16MS096)) 16MS096_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

Post-transcriptional gene expression regulation is influenced by crosstalk in between the vital regulatory molecules like RNA-binding proteins (RBPs) and non-coding RNAs (ncRNAs) both of which mostly interact with the 3’untranslated regions (3’UTRs) of mRNAs. Several studies have investigated the interplay between different microRNAs and RNA binding proteins (RBPs) in regulating target gene expression. Now in this above scenario, 6-adenosine methylation (m6A), the most prevalent and abundant internal modification in mRNAs of eukaryotes, adds a new dimension. m6A is found to be enriched in 3’UTRs of mRNAs. mRNA 3’untranslated regions are also the crucial targeting sites of miRNA and RBP, both of which are close to the m6A sites in HEK293/T cell lines. This suggests the possibility that m6A modification might affect the interplay between RBPs and miRNAs in binding to mRNAs. In this study, we have done a bioinformatic analysis to check whether there is a correlation between sites of m6A and the targeting sites RBP and miRNA in mRNAs. This study reveals that this correlation exists for the HeLa cell line and is cell line independent. We have further selected a gene, COX7A2L, which contains predicted binding sites of HuR and miR-125b in the 3’ untranslated region and is also reported to contain m6A sites nearby. We have used this gene to conduct preliminary experiments to investigate the possible influence of m6A on HuR and miR-125 binding as a “proof of concept” of the three-way crosstalk between miRNAs, RBPs, and mRNA modifications.

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