The picornaviruses are a family of small positive sense single stranded RNA viruses that cause a wide range of diseases at an annual cost well into the hundreds of million dollars. Members include acute hepatitis A virus, the heart disease-causing coxsackie B3 virus, rhinoviruses that cause more than half the occurrences of the common cold, and the paralyzing poliovirus. These viruses share a common life cycle where their RNA replication and viral assembly occurs in large membrane anchored replication complexes assembled on the surfaces of vesicles derived from the endoplasmic reticulum. The replication process is driven by a virally encoded RNA dependent RNA polymerase, the 3Dpol protein, that is responsible the synthesis of all viral RNA. Like all picornaviral proteins, the polymerase is generated by proteolytic cleavage of a single large viral polyprotein. There is mounting evidence in several picornaviruses that the polymerase and its immediate precursors are directly responsible for the assembly of these replication centers. The 3Dpol polymerase of poliovirus, the best studied of the picornaviruses, has been shown to assemble into large sheet structures along a protein-protein interface that was initially identified in a partial crystal structure of 3Dpol. We have solved the complete crystal structure of 3Dpol at 2.0 A resolution and discovered that the enzyme requires a free N-terminus to properly fold the active site, providing a molecular basis for the processing dependent activation of the polymerase. We are continuing our structural studies of the poliovirus proteins by further characterizing the conformational flexibility of 3Dpol and expanding into determining the role of protein-protein interfaces in the assembly of 3CDpro and other precursor proteins. The results will yield fundamental insights into the replication of this important group of pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI059130-04
Application #
7195791
Study Section
Virology Study Section (VR)
Program Officer
Park, Eun-Chung
Project Start
2004-04-01
Project End
2009-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
4
Fiscal Year
2007
Total Cost
$238,318
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
785979618
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
Rai, Devendra K; Diaz-San Segundo, Fayna; Campagnola, Grace et al. (2017) Attenuation of Foot-and-Mouth Disease Virus by Engineered Viral Polymerase Fidelity. J Virol 91:
Peersen, Olve B (2017) Picornaviral polymerase structure, function, and fidelity modulation. Virus Res 234:4-20
Karr, Jonathan P; Peersen, Olve B (2016) ATP Is an Allosteric Inhibitor of Coxsackievirus B3 Polymerase. Biochemistry 55:3995-4002
McDonald, Seth; Block, Andrew; Beaucourt, Stéphanie et al. (2016) Design of a Genetically Stable High Fidelity Coxsackievirus B3 Polymerase That Attenuates Virus Growth in Vivo. J Biol Chem 291:13999-4011
Sexton, Nicole R; Smith, Everett Clinton; Blanc, Hervé et al. (2016) Homology-Based Identification of a Mutation in the Coronavirus RNA-Dependent RNA Polymerase That Confers Resistance to Multiple Mutagens. J Virol 90:7415-7428
Kempf, Brian J; Peersen, Olve B; Barton, David J (2016) Poliovirus Polymerase Leu420 Facilitates RNA Recombination and Ribavirin Resistance. J Virol 90:8410-21
Svensen, Nina; Peersen, Olve B; Jaffrey, Samie R (2016) Peptide Synthesis on a Next-Generation DNA Sequencing Platform. Chembiochem 17:1628-35
Campagnola, Grace; McDonald, Seth; Beaucourt, Stéphanie et al. (2015) Structure-function relationships underlying the replication fidelity of viral RNA-dependent RNA polymerases. J Virol 89:275-86
Kao, C Cheng; Peersen, Olve B (2014) Editorial overview: Virus replication in animals and plants. Curr Opin Virol 9:iv-v
Sholders, Aaron J; Peersen, Olve B (2014) Distinct conformations of a putative translocation element in poliovirus polymerase. J Mol Biol 426:1407-19

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