Use of antivirals to probe flavivirus replication

Carolyn G Botting, Purdue University

Abstract

The flaviviruses, including yellow fever, dengue, and West Nile viruses are pathogens that affect millions of people each year. Despite the wide prevalence of flavivirus infections, there are no treatment options after infection occurs. A high-throughput study aimed at developing new flavivirus therapeutics previously identified two candidate antiviral compounds, 3394 and 4088, that proved to target yellow fever nonstructural protein 4B (NS4B). NS4B's role in infection is poorly understood, and thus it was of interest to better understand NS4B's function in infection as well as develop the compounds as antivirals. To this end, a variety of biochemical techniques were employed to determine whether the compounds target any of the known functions of NS4B. Specifically, it was determined that the compounds do not inhibit NS4B oligomerization, its interaction with NS3 or NS5, its immune response evasion activity, or its targeting to the endoplasmic reticulum membrane. Immunofluorescence imaging, qRT-PCR, and synergy assays with C75 (a fatty acid synthase inhibitor) proved that the compounds target some function of NS4B necessary in early replication complex formation, suggesting a function of NS4B that has not yet been characterized. Furthermore, second generation compounds with reduced cytotoxicity target NS4B in a similar manner, thus proving that these compounds are excellent therapeutic candidates and can be used to gain insight into NS4B's role in infection.

Degree

M.S.

Advisors

Kuhn, Purdue University.

Subject Area

Virology

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