HUMAN RNA METHYLTRANSFERASE METTL16 AFFECTS MULTIPLE CELL PROCESSES THROUGH DIFFERENTIAL FUNCTION OF ITS DOMAINS

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Emily Satterwhite Talic (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: ABSTRACT\n In recent years, disease therapies have been developed to target specific RNA and protein expression changes in the affected cells/tissue. The RNA post-transcriptional modification, methyl-6-adenosine (m6A), has been implicated in a number of disease-states as a driver of aberrant RNA and protein expression. m6A is the reversible methylation of adenosine which is produced by enzymes known as m6A RNA methyltransferases. There are currently two known m6A RNA methyltransferases which methylate messenger RNA in humans: METTL3/14 complex, and METTL16. The METTL3/14 complex has been widely implicated and studied in cancer development and progression, while much of METTL16 has not been investigated. While a few studies have probed the biochemical details of METTL16, cancer studies have shown METTL16 protein expression level changes have been associated with a variety of cancer types. The research in this dissertation project was performed in an effort to determine the RNAs affected by and the cell-wide effects of METTL16’s function. Using RNA immunoprecipitation, the studies here show that METTL16 binds a number of RNAs outside the 3 that are currently accepted as RNA targets. It was also discovered that METTL16 is localized to both the nucleus and cytoplasm, suggesting a role in addition to methylation in the cell. Mutation of key residues in METTL16 protein resulted in a large number of changes in RNA and protein expression levels determined with RNA sequencing, real-time PCR, and mass spectrometry proteomics. A majority of these RNAs and proteins are associated with cytoskeletal maintenance, calcium signaling, and cell cycle regulation, all of which have implications in distinct diseases including cancer. Of interest, only some of the METTL16 mutations seemed to affect cell cycle phase occupancy. Given the multiple changes seen among cells containing mutated METTL16, it is plausible that this protein could aid in presentation or progression of associated diseases and could therefore be a justified target for cell therapies.

Additional Information

Publication

Dissertation

Language: English

Date: 2023

Subjects

METTL16;RNA;m6A;methyltransferase;cancer

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