Cellular Metabolism and its Impact on Viral Infection
Abstract
Description
The prevalence of viruses and the impact they can have, as seen by the COVID-19 pandemic, clearly demonstrates the need to identify the mechanisms of various cellular innate antiviral defense pathways. One specific pathway that will be focused on in this research is the 2′-5′ oligoadenylate synthetase (OAS) / RNase L pathway, which is dependent on the amounts of cellular ATP for which I hypothesize that if cells are exposed to dsRNA or a viral infection, the activation of OAS will decrease cellular ATP level by converting ATP to 2-5A, providing an essential link of the metabolic processes during viral infection. In order to understand the relationship between the OAS/RNase L mechanism and cellular ATP, the procedures I will be utilizing for this experiment include cell culture, viral transfection using plasmid, luminescent assay, RNA gel electrophoresis, Western blot, and fluorescence assay. These tools will be used to maintain stable cell lines expressing the proper knockout genes, introduce viral RNA to the cells, measure initial and final amounts of ATP, observe the degradation of RNA and inhibit other defense pathways from being activated. Based on literature and prior research, it is expected that viral transfections will lower the ATP levels in the cells while also causing the degradation of dsRNA. The data collected from gel electrophoresis clearly indicates that upon viral transfection, the dsRNA in the cell is degraded due to the activation of the RNase L. Moreover, it is predicted that the three OAS gene knockouts will have varying impact on the response to viral RNA based on the amount of ATP consumed to create 2-5As. This project can yield a great deal of information regarding innate immunity that can be used in both clinical and pharmaceutical settings, drastically improving the quality of life for many individuals. Moreover, this research will pose new questions regarding how cells resist viral infections due to ! other stressors, various plasmids, and the introduction of different viral proteins.
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Keywords
OAS, RNase L, ATP, Virus