Date of Award

1-1-2021

Language

English

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

College/School/Department

Department of Biological Sciences

Content Description

1 online resource (xvii, 192 pages) : illustrations.

Dissertation/Thesis Chair

Cara T Pager

Committee Members

Daniele Fabris, Alexander T Ciota, Gabriele Fuchs

Keywords

epitranscriptomics, Hepatitis C virus, mass spectrometry, methylation, RNA modification, Zika virus, RNA, Transcription factors, Mosquitoes as carriers of disease, Mass spectrometry, RNA editing, Virus diseases, Viral genetics

Subject Categories

Virology

Abstract

Viral epitranscriptomics is a novel field of RNA biology. Chemical groups are appended to RNA nucleotides under different conditions in cells, and I examine these changes using a mass spectrometry-based approach. Various changes are found to occur in post-transcriptional modifications (PTMs) during viral infection. The field primarily studies a small number of PTMS, such as m6A, while I explore a variety of novel modifications and their impact on viral infection. Hepatitis C virus (HCV) infection induces a variety of modification changes, including the novel presence of dimethylcytosine (m5Cm/m44C). One Writer enzyme for this modification is determined to likely be METTL15, where knockdown of this enzyme causes PTM-level changes and a dramatic increase in HCV protein and RNA levels. During ZIKV infection, PTM changes are induced in numerous mammalian cell lines, as well as in mosquito cell lines. I then perform infections in whole in vivo mosquitoes and note dramatic increases in dimethylguanosine and methyladenosine. These modifications are sufficient to separate the Mock-infected mosquitoes from the ZIKV-infected mosquitoes and could lead to a means of viral detection in unknown mosquito samples. Future work will investigate novel Writer proteins, purpose of viral PTMs, and viral genome PTM changes occurring between host organisms.

Available for download on Thursday, May 22, 2025

Included in

Virology Commons

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