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Journal of Virology, May 2005, p. 6172-6179, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6172-6179.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Polypyrimidine Tract Binding Protein Is Required for Efficient Picornavirus Gene Expression and Propagation

Paola M. Florez,1,{dagger} October M. Sessions,1,{dagger} Eric J. Wagner,1,2,{dagger},{ddagger} Matthias Gromeier,1,3* and Mariano A. Garcia-Blanco1,3*

Department of Molecular Genetics and Microbiology,1 Program in Molecular Cancer Biology,2 Department of Medicine, Duke University Medical Center, Durham, North Carolina 277103

Received 11 June 2004/ Accepted 12 January 2005

Mammalian host factors required for efficient viral gene expression and propagation have been often recalcitrant to genetic analysis. A case in point is the function of cellular factors that trans-activate internal ribosomal entry site (IRES)-driven translation, which is operative in many positive-stranded RNA viruses, including all picornaviruses. These IRES trans-acting factors have been elegantly studied in vitro, but their in vivo importance for viral gene expression and propagation has not been widely confirmed experimentally. Here we use RNA interference to deplete mammalian cells of one such factor, the polypyrimidine tract binding protein, and test its requirement in picornavirus gene expression and propagation. Depletion of the polypyrimidine tract binding protein resulted in a marked delay of particle propagation and significantly decreased synthesis and accumulation of viral proteins of poliovirus and encephalomyocarditis virus. These effects could be partially restored by expression of an RNA interference-resistant exogenous polypyrimidine tract binding protein. These data indicate a critical role for the polypyrimidine tract binding protein in picornavirus gene expression and strongly suggest a requirement for efficient IRES-dependent translation.

* Corresponding author. Mailing address for M. Gromeier: Duke University Medical Center, Box 3020 (451 Jones), Research Drive, Durham, NC 27710. Phone: (919) 668-6205. Fax: (919) 684-8735. E-mail: grome001{at}mc.duke.edu. Mailing address for M. A. Garcia-Blanco: Duke University Medical Center, Box 3053 (424 CARL), Research Drive, Durham, NC 27710. Phone: (919) 613-8632. Fax: (919) 613-8646. E-mail: garci001{at}mc.duke.edu.

{dagger} Paola M. Florez, October M. Sessions, and Eric J. Wagner contributed equally to this work.

{ddagger} Present address: Department of Biochemistry and Biophysics, Program in Molecular Biology and Biotechnology, CB#7100, University of North Carolina, Chapel Hill, NC 27599.

Journal of Virology, May 2005, p. 6172-6179, Vol. 79, No. 10
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.10.6172-6179.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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