This proposal describes experiments that are concerned with the study of poliovirus replication and pathogenesis. An infectious viral cDNA clone will be mutagenized to create a collection of altered plasmids which will be used for most of the proposed studies. The functions of the central (P2) region of the genome and the 5'- and 3' -untranslated sequences will be addressed by studying viable virus mutants containing alterations in these regions. Mutated, noninfectious cDNAs which contain lesions in these areas of the genome will be studied by synthesizing positive strand RNA and transfecting this RNA into cultured cells. The role of terminal nucleotide sequences of the viral RNA in translation and replication will be studied using in vitro systems employing as templates mutated, subgenomic RNA fragments. Questions on the biology of poliovirus defective-interfering particles will be addressed. Positive strand RNAs will be synthesized which contain deletions in the capsid region, and these RNAs will be transfected into cells to determine whether DI RNAs can replicate, persist in and be rescued from mammalian cells. A mouse model for poliomyelitis will be studied to identify viral functions which participate in the production of paralytic disease. Mouse-avirulent viral mutants will be isolated and used to study the molecular basis of neurovirulence. The mouse model will also be used to identify viral capsid protein sequences required for entry of poliovirus into cells of the mouse central nervous system. To better understand how the viral capsid interacts with a cell during initiation of an infectious cycle, the cellular receptor for poliovirus will be studied. DNA transfection techniques will be used to isolate molecular clones of the cellular gene encoding the poliovirus receptor. These studies are part of our long-term research goal, to provide a complete description of the replication of a human pathogen.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI020017-04
Application #
3129494
Study Section
Experimental Virology Study Section (EVR)
Project Start
1983-04-01
Project End
1988-03-31
Budget Start
1986-06-01
Budget End
1987-05-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Kim, Melissa Stewart; Racaniello, Vincent R (2007) Enterovirus 70 receptor utilization is controlled by capsid residues that also regulate host range and cytopathogenicity. J Virol 81:8648-55
Kauder, Steven E; Racaniello, Vincent R (2004) Poliovirus tropism and attenuation are determined after internal ribosome entry. J Clin Invest 113:1743-53
Brown, David M; Kauder, Steven E; Cornell, Christopher T et al. (2004) Cell-dependent role for the poliovirus 3' noncoding region in positive-strand RNA synthesis. J Virol 78:1344-51
Tsang, S K; McDermott, B M; Racaniello, V R et al. (2001) Kinetic analysis of the effect of poliovirus receptor on viral uncoating: the receptor as a catalyst. J Virol 75:4984-9
McDermott Jr, B M; Rux, A H; Eisenberg, R J et al. (2000) Two distinct binding affinities of poliovirus for its cellular receptor. J Biol Chem 275:23089-96
Dove, A W; Racaniello, V R (2000) An antiviral compound that blocks structural transitions of poliovirus prevents receptor binding at low temperatures. J Virol 74:3929-31
Belnap, D M; McDermott Jr, B M; Filman, D J et al. (2000) Three-dimensional structure of poliovirus receptor bound to poliovirus. Proc Natl Acad Sci U S A 97:73-8
Zhang, S; Racaniello, V R (1997) Persistent echovirus infection of mouse cells expressing the viral receptor VLA-2. Virology 235:293-301
Dove, A W; Racaniello, V R (1997) Cold-adapted poliovirus mutants bypass a postentry replication block. J Virol 71:4728-35
Liao, S; Racaniello, V (1997) Allele-specific adaptation of poliovirus VP1 B-C loop variants to mutant cell receptors. J Virol 71:9770-7

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