Adenosine triphosphate is an allosteric inhibitor of coxsackievirus B3 3Dpol

Picornaviruses pose a significant threat to human and animal health, but at present there are no drugs to prevent or treat picornaviral infections. However, intensive study of the picornaviral lifecycle has revealed several promising pharmacological targets, including the RNA-dependent RNA polymerase, 3Dpol, that is responsible for replicating the viral genome. 3Dpol is central in the virus lifecycle, determines the distribution of mutants in viral progeny, and is a very highly conserved protein across picornavirus species. As such, it is an attractive target for antiviral research. The only 3Dpol inhibitors that have been found to date are nucleoside analogues that act directly on the active site, even though global dynamics of the protein that are sensitive to allosteric effects of various mutations have been shown to be important determinants of fidelity. The research presented in this thesis provides the first direct evidence of allosteric regulation of 3Dpol by a small molecule. Inhibition assays investigating the relative affinities of a stalled coxsackievirus elongation complex for non-cognate nucleotides uncovered a mixed inhibition profile of ATP. Among the six picornavirus species tested, this mode of inhibition seems specific to coxsackievirus B3 (CVB3). Engineered mutations in CVB3 3Dpol, including two that were previously found to lower polymerase fidelity, diminish the uncompetitive component of ATP inhibition. ATP inhibition was found to be dependent on the β- and γ-phosphates. The potential role of ATP’s allosteric effect in the virus lifecycle as well as the importance of a biochemically confirmed allosteric site on the polymerase are discussed.

Name thesis:degree_name

Master of Science (M.S.)

Level thesis:degree_level

Masters

Discipline thesis:degree_discipline

Biochemistry and Molecular Biology

Grantor dc:publisher

Colorado State University. Libraries

Year dc:date.issued

2016

Authors dc:creator

• Karr, Jonathan Paul, author
• Peersen, Olve, advisor
• Cohen, Robert, committee member
• Perera, Rushika, committee member

• ATP
• polymerase
• allostery
• uncompetitive inhibition
• Coxsackievirus B3

Statement dc:rights

• Copyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.

Language dc:language.iso

eng, English

Identifier URI

https://doi.org/10.25675/3.018403

OAI identifier oai:identifier

oai:mountainscholar.org:10217/173545