Date of Award

January 2021

Document Type

Open Access Thesis

Degree Name

Master of Public Health (MPH)

Department

School of Public Health

First Advisor

Nancy Ruddle

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of acute respiratory coronavirus disease 2019 (COVID-19), a disease that erupted into a global pandemic in March of 2020.2 Recently, a genome wide CRISPR screen was performed on SARS-lineage coronaviruses and two additional recombinant viruses; the screen revealed host genes essential to coronavirus pathogenesis and entry.7 The screen identified KDM6A, a histone demethylase, as an important pro-viral gene.7 Building upon this observation, in this thesis, I explore the role of Kdm6a in murine hepatitis virus (MHV), a type of mouse coronavirus. MHV is the prototypical model for studying SARS-CoV-2 pathogenesis due to the ease of biological safety protocols and its ability to help us characterize coronaviruses generally.41 The Kdm6a gene encodes enzyme lysine specific demethylase 6A that is found in many cells of the body.8 Kdm6a functions as a histone demethylase, which helps increase activity of certain genes, specifically developmental genes.8 Though much is not known about Kdm6a’s implication in coronavirus infection, preliminary data suggests that Kdm6a is essential to MHV-A59 and MHV-3 infection. In our first aim, we performed RT-qPCR to elucidate Kdm6a’s role in viral entry. We found Kdm6a knockout results in reduced Ceacam1 expression, the receptor for MHV. For our second aim, we designed mutated Kdm6a constructs to assess which domains of Kdm6a are important in viral infection. We proceeded to run rescue assays to observe expression of Kdm6a in Kdm6a deficient cells. Though we did not observe expression, there are likely explanations as to why these experiments did not work. To this end, the present study suggests that Kdm6a is important in viral entry, possibly through regulation of receptor expression, but which of its domains are responsible remains to be elucidated.

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This is an Open Access Thesis.

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