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Glycolytic control of vacuolar ATPase pump activity: a mechanism to regulate influenza viral infection

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Hinissan Kohio (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Advisor
Amy Adamson

Abstract: Influenza viruses are pathogens that can cause major pandemics in the human population, such as the Spanish Flu outbreak in 1918-1919, which caused about 50 million deaths worldwide. As the virus frequently mutates, vaccines and anti-viral drugs can be rendered ineffective over time. As new influenza viruses emerge, new anti-viral therapies are essential. The goal of this study was to identify novel means to inhibit influenza viral replication, by targeting the activity of the Vacuolar ATPase (V-ATPase) proton pump. The V-ATPase pump is a multi-molecular proton pump that couples ATP hydrolysis to proton transport, thus regulating the pH of intracellular compartments. This activity is also used by the influenza virus to induce viral uncoating during infection. In this study, I examined how changes in glucose concentrations affect V-ATPase pump activity and influenza replication. Through immunostaining of mammalian cells, I observed that higher levels of glucose caused specific localization of the V-ATPase proton pumps within the cell, thus triggering pump activity. Western blot analysis of two V-ATPase proton pump proteins showed that protein synthesis of the pump was not affected by increasing the amount of glucose. Based on these results, I examined the effect of increasing glucose levels upon influenza infection. I found that higher amounts of glucose yielded higher influenza infection levels. Additionally, viral infection was significantly reduced after inhibition of glycolysis with 2-deoxyglucose and 3-bromopyruvate. Lastly, I bypassed the need for glycolysis via addition of extracellular ATP. The V-ATPase pump activity was restored after glycolytic inhibition with ATP treatment as indicated by an increased number of virally infected cells. Taken together, I propose that inhibiting influenza replication in mammalian cell cultures by altering activity of the V-ATPase proton pump via inhibition of glycolysis could be a potential new approach for the treatment of influenza infection.

Additional Information

Publication
Thesis
Language: English
Date: 2012
Keywords
Glycolysis, Influenza, V-ATPase
Subjects
Influenza $x Research