Structural Analysis of Virulence Determinants and Genomic DNA Packaging Constraints in a ssDNA Virus

University Of Florida, Gainesville FL

Investigators

Abstract

The three-dimensional (3D) structure of a macromolecule is assembled to enable functional utilization. This 3D structure, however, once conformed, is capable of transitions and/or re-arrangements, as required, to enable essential functional interactions. The consecutive macromolecular interactions that define the life-cycle of viruses during host-cell infection represent excellent examples of the high fidelity of functional utilization of macromolecular structure. However, despite much research on viral life cycles, there is limited 3D structural information on the facilitating capsid-host interactions, structural capsid transitions, and capsid-genome interactions. This project will utilize naturally-occurring variants and emerging virulent strains of the Minute Virus of Mice (MVM), a member of the ssDNA Parvoviridae, as a viral model to decipher the structural requirements for successful host-cell receptor recognition, cellular trafficking and capsid maturation triggered by genomic interaction. This model system is also ideal for studies of virus strains that are similar to one another yet have pronounced differences in their host-range, tissue tropism, and pathogenic outcomes of infection due to minor structural variations or amino-acid differences on their capsid viral proteins which modify required infectious interactions. These variations often confer altered viral host-cell interactions, including infectious receptor utilization, which is common in emerging viruses. This project includes a multi-disciplinary approach, utilizing X-ray crystallography, biochemistry, biophysics, and virology, in a collaborative effort among investigators from four different institutions. Research outcomes will augment the understanding of functional domain utilization for viral infection in general, particularly for non-enveloped viruses that must evolve strategies for traversing the complex cellular milieu en route to and from their sites of replication, and provide further insight into the mechanism(s) of viral adaptations to new hosts, a phenomenon that often gives rise to emerging viral pathogens. Broader Impacts: The research objectives of this project will be integrated into the educational training of high school, undergraduate and graduate students in the use of a multidisciplinary approach for studies aimed at deciphering structure-function correlations for macromolecular interactions. Specifically, high school students will be recruited to work on this project during the summer months through the University of Florida Student Science Training Program (SSTP), three undergraduate students are already actively involved in this project, and three of the specific research aims will form the research component of the Ph.D thesis of a graduate student. Undergraduate and graduate students will also participate in the research studies to be conducted in the collaborating laboratories. In addition, a specific outreach aim will involve the production of an educational CD-ROM titled "Structural biology as a tool for scientific research" as part of the University of Florida "Excursion in Science" Program web based learning tools aimed at K-12 students. This aim will also include the development of a three day workshop as part of the University of Florida "Teachers as Scholars" summer program to accompany the CD-ROM that will provide interdisciplinary physics, math, and biology high school teachers with hands-on experience and theoretical knowledge of structural biology that can be integrated into their classroom teaching strategies.