The investigation of PRMT5 in cancer through biochemical and computational approaches.
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Date
2023-12
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The Ohio State University
Abstract
The protein-arginine methyltransferases (PRMT) family of proteins function to methylate numerous nuclear and cytoplasmic substrates and play a critical role in the regulation of important biological processes such as cell cycle progression, cell apoptosis, RNA processing, and DNA damage repair. PRMT's are also by definition epigenetic regulators due to their ability to participate in chromatin remodeling by way of histone post-translational modification. There are several PRMT's found in humans and PRMT5 specifically has been shown to play a key role in the development and progression of multiple forms of cancer. Due to PRMT5's significant role in cancer, we profiled cancer-associated mutations and investigated a novel approach to inhibit its activity. In this thesis, a computational genomic and structural biochemistry approach was used to analyze PRMT5 cancer-associated mutation identified in the Catalogue of Somatic Mutations in Cancer (COSMIC). Then, the yeast homolog Skb1 was expressed and purified to perform site-directed mutagenesis and methyltransferase activity assays. The computational mutation study on PRMT5 yielded the discovery of 3 driver mutations and several pathogenic mutations. The yeast analog of PRMT5, Skb1, was purified as a recombinant protein and a fluorescent methyltransferase assay was used to determine biochemical activity.