C36: Virus hopping between animal and human: Structural characterization of equine G14P[12] rotavirus via X-ray crystallography

Rotavirus is a virus that infects both humans and animals. Group A Rotaviruses are comprised of five genogroups P[1] through P[V], which are further organized into 37 genotypes P[1] through P[37]. These categorizations are based on sequence comparisons of spike proteins VP4 and VP7. As a general rule, P[I] RV’s infect animals, and P[II] infect humans. However, some members of genogroups P[I], P[II], P[IV], and particularly P[III] are able to infect both, indicating the presence of mechanistic commonalities among RV variants resulting from mutations affecting the VP4 and VP7 spike proteins, which are the proteins responsible for binding to host receptor cells. The combination of VP7 and VP4 proteins present on any given variant affect its binding affinity and specificity. Comparison of binding characteristics among variants provide insight into RV zoonosis. Here, G12P[14], an equine rotavirus, is overexpressed, purified, and studied via X-ray crystallography; this involves the use of analytical techniques common to biochemistry and structural biology such as a nickel affinity column, Bradford assay, SDS-PAGE with Coomassie blue stain, hanging-drop vapor-diffusion crystallization, and X-ray crystallography. Diffraction data were collected at the Advanced Photon Source (APS) beamline at the Argonne National Laboratory in Chicago, IL. The phylogenetic observations made here may apply to broader topics within virology and epidemiology such as Covid-19 with regard to zoonosis. Future work may focus on solving the crystal structure in complex with ligand and determining this variant’s binding affinity with ITC or HSQC-NMR.

Author(s): Meredith Hansen, Biochemistry Major

Advisor(s): Michael Kennedy, Department of Chemistry and Biochemistry