Investigating Regions of Interest in Chikungunya Virus Nonstructural Proteins (2023)

Undergraduates: Ian Klevans, Jennifer Loome

Faculty Advisor: Mark Heise
Department: Genetics, Biology

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Chikungunya virus is a member of the Alphavirus genus that infects humans and may cause severe illness. The viral genome encodes viral proteins, one of which is nonstructural protein 2 (nsp2). One of the functions of nsp2 is its ability to shut off cellular transcription via the degradation of RNA Polymerase Binding Domain 1, or RPB1. Shutoff of cellular transcription is cytopathic, or cell-killing. This experiment explored which regions in nsp2 are necessary for this cytopathic effect. Here we mutated specific amino acid regions of interest within nsp2, created a pCMV plasmid vector system to express mutant nsp2, transfected these mutant nsp2 plasmids into cells, and observed cell death. In future experiments, we hope to co-express a fluorescent tagged RPB1 with nsp2, allowing for more specific determination of which mutants lost the ability to degrade RPB1 after transfection.

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