The project described in this proposal will study the processing of poliovirus proteins during a viral infection. A molecular genetic approach with infectious cDNA clones of the viral RNA as targets for site-specific mutagenesis will be used to define the specific regions of the viral genome involved in the enzyme-substrate interactions of proteolytic processing. A series of defined, mutant polio plasmids will be analyzed for phenotypic and biochemical expression in the following four different assay systems: (1) transfection of eukaryotic cells with polio plasmids, (2) rescue of proteinase mutants of infectious recombinant plasmids by co-transfection with simian virus 40 (SV40) vectors that express the P3 (replicase) region polypeptides of poliovirus, (3) bacterial expression of specific poliovirus polypeptides, and (4) in vitro translation of mutant mRNAs derived from bacteriophage SP6-specific transcription of polio cDNA. The proposed experiments will define the functional domains of the viral proteinase as well as those regions of polio proteins that are important in the formation of authentic substrate polypeptides. The data obtained should provide mechanisms for how the regulation of cleavage of poliovirus precursor polypeptides by a small genomic RNA can control both viral and cellular biosynthetic activity. Such a unique regulation of gene expression must contribute to the effectiveness of poliovirus and the other picornaviruses as cellular pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI022693-01
Application #
3134161
Study Section
Virology Study Section (VR)
Project Start
1985-07-01
Project End
1988-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
Schools of Medicine
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697
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Hopcraft, Sharon E; Azarm, Kristopher D; Israelow, Benjamin et al. (2016) Viral Determinants of miR-122-Independent Hepatitis C Virus Replication. mSphere 1:
Ullmer, Wendy; Semler, Bert L (2016) Diverse Strategies Used by Picornaviruses to Escape Host RNA Decay Pathways. Viruses 8:
Wehbe, Michel; Huguenin, Antoine; Leveque, Nicolas et al. (2016) Construction of a subgenomic CV-B3 replicon expressing emerald green fluorescent protein to assess viral replication of a cardiotropic enterovirus strain in cultured human cells. J Virol Methods 230:1-8
Flather, Dylan; Cathcart, Andrea L; Cruz, Casey et al. (2016) Generation of Recombinant Polioviruses Harboring RNA Affinity Tags in the 5' and 3' Noncoding Regions of Genomic RNAs. Viruses 8:
Flather, Dylan; Semler, Bert L (2015) Picornaviruses and nuclear functions: targeting a cellular compartment distinct from the replication site of a positive-strand RNA virus. Front Microbiol 6:594
Lévêque, Nicolas; Semler, Bert L (2015) A 21st century perspective of poliovirus replication. PLoS Pathog 11:e1004825
Ertel, Kenneth J; Brunner, Jo Ellen; Semler, Bert L (2010) Mechanistic consequences of hnRNP C binding to both RNA termini of poliovirus negative-strand RNA intermediates. J Virol 84:4229-42
Brunner, Jo Ellen; Ertel, Kenneth J; Rozovics, Janet M et al. (2010) Delayed kinetics of poliovirus RNA synthesis in a human cell line with reduced levels of hnRNP C proteins. Virology 400:240-7
Sean, P; Semler, B L (2008) Coxsackievirus B RNA replication: lessons from poliovirus. Curr Top Microbiol Immunol 323:89-121

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