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 life cycle where 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 for the synthesis of all viral RNA. This research project is focused on the structure and assembly of viral replication centers, where we use poliovirus and coxsackievirus as our main experimental systems. We have previously solved the crystal structures of the 3Dpol proteins from both these viruses and elucidated the molecular mechanism behind the proteolytic activation of these proteins upon cleavage from the viral 3CDpro precursor protein. We are continuing our studies of picornaviral replication proteins by focusing on the structural changes associated with the formation of the 3Dpol elongation complex and on understanding how mutations in the polymerase affect viral RNA replication rate and fidelity both in vitro and in vivo. A new aspect of the project is focused on the structure and function of the membrane associated viral 2C and 2BC proteins that are responsible for host cell membrane rearrangements resulting in the formation of the vesicles upon which the viral replication complexes assemble.

Public Health Relevance

Poliovirus is a member of a large family of viruses containing a specific RNA dependent RNA polymerase protein that is responsible for replicating the viral genome. The focus of this research project is to understand how this protein functions during virus replication and find ways to interfere with its function, which can open the door to the development of antiviral drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI059130-10
Application #
8658796
Study Section
Virology - A Study Section (VIRA)
Program Officer
Park, Eun-Chung
Project Start
2004-04-01
Project End
2015-05-31
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
10
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
City
Fort Collins
State
CO
Country
United States
Zip Code
80521
Peersen, Olve B (2017) Picornaviral polymerase structure, function, and fidelity modulation. Virus Res 234:4-20
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:
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 90:7415-28
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
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
Sholders, Aaron J; Peersen, Olve B (2014) Distinct conformations of a putative translocation element in poliovirus polymerase. J Mol Biol 426:1407-19
Kao, C Cheng; Peersen, Olve B (2014) Editorial overview: Virus replication in animals and plants. Curr Opin Virol 9:iv-v

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