The purpose of this project is to understand the molecular mechanisms responsible for replication of picornaviruses in susceptible target cells. This virus family includes numerous human pathogens (poliovirus, coxsackievirus, echovirus, enteroviruses, rhinoviruses, hepatitis A virus). Infection of cells with these viruses leads to major changes in the host cell's structure and metabolic activity. Cellular protein and RNA synthesis are inhibited; the intracellular membrane network becomes rearranged into vesicles that surround and provide a scaffold for viral RNA replication complexes; cellular proteins are subverted into facilitating viral protein and RNA synthesis. The unique combination of viral and cellular proteins together accomplish a highly efficient production of viral RNA, proteins, and particles. We are studying the activities of individual viral gene products, expressed alone or in combination, in cultured human cells, to determine their specific roles in the replication process. Assays for individual steps in the reaction have been developed and are being characterized biochemically. Mutations in individual viral proteins have been engineered and analyzed for their effects on different steps in the replication process. We have discovered that synthesis of specific viral protein(s) induce tha activation and recruitment to membranes of a group of cellular proteins that play specific roles in membrane trafficking, vesicle formation, and morphology, which collectively results in the formation of viral RNA replication complexes in infected cells. these cellyular proteins appear to trigger the formation of a membranous scaffold for viral RNA synthesis, and perhaps recruit additional proteins that are required for virus replication. Understanding the precise biochemical activities of viral proteins in the replication process will allow the development of new strategies for vaccine development and the design of antiviral drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000816-07
Application #
6986585
Study Section
(LID)
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2004
Total Cost
Indirect Cost
Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Belov, George A; Habbersett, Courtney; Franco, David et al. (2007) Activation of cellular Arf GTPases by poliovirus protein 3CD correlates with virus replication. J Virol 81:9259-67
Belov, George A; Altan-Bonnet, Nihal; Kovtunovych, Gennadiy et al. (2007) Hijacking components of the cellular secretory pathway for replication of poliovirus RNA. J Virol 81:558-67
Belov, George A; Ehrenfeld, Ellie (2007) Involvement of cellular membrane traffic proteins in poliovirus replication. Cell Cycle 6:36-8
Chumakov, Konstantin; Ehrenfeld, Ellie; Wimmer, Eckard et al. (2007) Vaccination against polio should not be stopped. Nat Rev Microbiol 5:952-8
Teterina, Natalya L; Gorbalenya, Alexander E; Egger, Denise et al. (2006) Testing the modularity of the N-terminal amphipathic helix conserved in picornavirus 2C proteins and hepatitis C NS5A protein. Virology 344:453-67
Teterina, Natalya L; Levenson, Eric; Rinaudo, Mario S et al. (2006) Evidence for functional protein interactions required for poliovirus RNA replication. J Virol 80:5327-37
Agol, Vadim I; Chumakov, Konstantin; Ehrenfeld, Ellie et al. (2005) Don't drop current vaccine until we have new ones. Nature 435:881
Belov, George A; Fogg, Mark H; Ehrenfeld, Ellie (2005) Poliovirus proteins induce membrane association of GTPase ADP-ribosylation factor. J Virol 79:7207-16
Harrison, Stephen C; Alberts, Bruce; Ehrenfeld, Ellie et al. (2004) Discovery of antivirals against smallpox. Proc Natl Acad Sci U S A 101:11178-92
Teterina, Natalya L; Rinaudo, Mario S; Ehrenfeld, Ellie (2003) Strand-specific RNA synthesis defects in a poliovirus with a mutation in protein 3A. J Virol 77:12679-91

Showing the most recent 10 out of 18 publications