This is an application for renewal of a grant to study picornavirus genome replication. Picornaviruses represent an existing and emerging threat to US public health. Although protein factors and genetic elements required for picornavirus genome replication are known and appear to be conserved, a clear understanding of the mechanisms employed to produce picornaviral RNA is lacking. The long-term objective of this program is to reconstitute picornavirus genome replication in vitro from purified components. We have achieved all of the major objectives of the previous funding period. In addition, we have solved the first crystal structure for a picornaviral 3CD protein, developed the technology to study 3C-RNA interactions by using nuclear magnetic resonance spectroscopy, and discovered that the 3CD protein has both pre- and post-replication functions. During the next funding period, we will continue our studies of picornavirus genome replication as well as explore our newly discovered function for 3CD by pursuing the following specific aims: (1) Define the mechanism of assembly and structural organization of the picornavirus VPg uridylylation complex by using molecular genetic, biochemical and biophysical approaches;(2) Define the molecular basis for sequence- and structure-specific RNA recognition by 3C by using nuclear magnetic resonance spectroscopy;and (3) Elucidate the function of 3CD in formation of replication complexes.
Picornaviruses represent an existing and emerging threat to US public health. Achievement of the goals of the application will provide novel targets and mechanisms for development of inhibitors to treat infections by picornaviruses, especially those for which vaccines are not available.
|Huang, Peng-Nien; Jheng, Jia-Rong; Arnold, Jamie J et al. (2017) UGGT1 enhances enterovirus 71 pathogenicity by promoting viral RNA synthesis and viral replication. PLoS Pathog 13:e1006375|
|Shengjuler, Djoshkun; Chan, Yan Mei; Sun, Simou et al. (2017) The RNA-Binding Site of Poliovirus 3C Protein Doubles as a Phosphoinositide-Binding Domain. Structure 25:1875-1886.e7|
|Shengjuler, Djoshkun; Sun, Simou; Cremer, Paul S et al. (2017) PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions. J Vis Exp :|
|Chan, Yan M; Moustafa, Ibrahim M; Arnold, Jamie J et al. (2016) Long-Range Communication between Different Functional Sites in the Picornaviral 3C Protein. Structure 24:509-517|
|Kolli, Swapna; Meng, Xiangzhi; Wu, Xiang et al. (2015) Structure-function analysis of vaccinia virus H7 protein reveals a novel phosphoinositide binding fold essential for poxvirus replication. J Virol 89:2209-19|
|Moustafa, Ibrahim M; Gohara, David W; Uchida, Akira et al. (2015) Conformational Ensemble of the Poliovirus 3CD Precursor Observed by MD Simulations and Confirmed by SAXS: A Strategy to Expand the Viral Proteome? Viruses 7:5962-86|
|Liu, Yen-Chin; Kuo, Rei-Lin; Lin, Jing-Yi et al. (2014) Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. PLoS Pathog 10:e1004199|
|Graci, Jason D; Gnadig, Nina F; Galarraga, Jessica E et al. (2012) Mutational robustness of an RNA virus influences sensitivity to lethal mutagenesis. J Virol 86:2869-73|
|Trahey, Meg; Oh, Hyung Suk; Cameron, Craig E et al. (2012) Poliovirus infection transiently increases COPII vesicle budding. J Virol 86:9675-82|
|Weeks, Spencer A; Lee, Cheri A; Zhao, Yan et al. (2012) A Polymerase mechanism-based strategy for viral attenuation and vaccine development. J Biol Chem 287:31618-22|
Showing the most recent 10 out of 23 publications