The long-range goal of this project is a mechanistic picture of viral entry, uncoating, replication, and assembly, derived from structure-based """"""""molecular movies"""""""" of these processes. A large part of our research during the coming five-year period will concentrate on the molecular rearrangements that accompany penetration across a cell membrane of non-enveloped viruses - in particular, reoviruses and rotaviruses. We propose X-ray crystallographic studies of viral outer-shell proteins (u1, the reovirus penetration protein; VP4, the rotavirus binding and penetration protein; VP7, the rotavirus glycoprotein), crystallographic studies of rotavirus inner capsid particles, and biochemical studies of the reovirus u1 conformational changes that accompany penetration. We will combine structures of proteins and protein complexes with images from electron cryomicroscopy of intact virus particles, to obtain """"""""hybrid"""""""" models of structures and intermediates that do not crystallize. We anticipate that studies of the rotavirus outer-shell proteins will facilitate design of recombinant vaccines. We will also expand our studies of the reovirus and rotavirus polymerase complexes. We can obtain a direct picture of transcription and replication, because multiple rounds of nucleotide addition can occur within crystals of the reovirus polymerase, lambda3. Finally, we will extend our structural studies of the human papillomavirus capsid protein to analyze full-sized virus-like particles, which are candidate immunogens in current vaccine trials.
| Salgado, Eric N; Garcia Rodriguez, Brian; Narayanaswamy, Nagarjun et al. (2018) Visualization of Calcium Ion Loss from Rotavirus during Cell Entry. J Virol 92: |
| Chao, Luke H; Jang, Jaebong; Johnson, Adam et al. (2018) How small-molecule inhibitors of dengue-virus infection interfere with viral membrane fusion. Elife 7: |
| Salgado, Eric N; Upadhyayula, Srigokul; Harrison, Stephen C (2017) Single-particle detection of transcription following rotavirus entry. J Virol : |
| Harrison, Stephen C (2017) Protein tentacles. J Struct Biol 200:244-247 |
| Kim, Irene S; Jenni, Simon; Stanifer, Megan L et al. (2017) Mechanism of membrane fusion induced by vesicular stomatitis virus G protein. Proc Natl Acad Sci U S A 114:E28-E36 |
| Harrison, Stephen C (2015) Viral membrane fusion. Virology 479-480:498-507 |
| Mahmutovic, Selma; Clark, Lars; Levis, Silvana C et al. (2015) Molecular Basis for Antibody-Mediated Neutralization of New World Hemorrhagic Fever Mammarenaviruses. Cell Host Microbe 18:705-13 |
| Abdelhakim, Aliaa H; Salgado, Eric N; Fu, Xiaofeng et al. (2014) Structural correlates of rotavirus cell entry. PLoS Pathog 10:e1004355 |
| Chao, Luke H; Klein, Daryl E; Schmidt, Aaron G et al. (2014) Sequential conformational rearrangements in flavivirus membrane fusion. Elife 3:e04389 |
| Estrozi, Leandro F; Settembre, Ethan C; Goret, Gaël et al. (2013) Location of the dsRNA-dependent polymerase, VP1, in rotavirus particles. J Mol Biol 425:124-32 |
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