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Journal of Virology, June 2006, p. 5327-5337, Vol. 80, No. 11
0022-538X/06/$08.00+0     doi:10.1128/JVI.02684-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evidence for Functional Protein Interactions Required for Poliovirus RNA Replication

Natalya L. Teterina,1 Eric Levenson,1 Mario S. Rinaudo,1 Denise Egger,2 Kurt Bienz,2 Alexander E. Gorbalenya,3 and Ellie Ehrenfeld1*

Laboratory of Infectious Diseases, NIAID, NIH, Bethesda, Maryland 20892,1 Institute for Medical Microbiology, University of Basel, Basel, Switzerland,2 Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands3

Received 21 December 2005/ Accepted 8 March 2006

Poliovirus protein 2C contains a predicted N-terminal amphipathic helix that mediates association of the protein with the membranes of the viral RNA replication complex. A chimeric virus that contains sequences encoding the 18-residue core from the orthologous amphipathic helix from human rhinovirus type 14 (HRV14) was constructed. The chimeric virus exhibited defects in viral RNA replication and produced minute plaques on HeLa cell monolayers. Large plaque variants that contained mutations within the 2C-encoding region were generated upon subsequent passage. However, the majority of viruses that emerged with improved growth properties contained no changes in the region encoding 2C. Sequence analysis and reconstruction of genomes with individual mutations revealed changes in 3A or 2B sequences that compensated for the HRV14 amphipathic helix in the polio 2C-containing proteins, implying functional interactions among these proteins during the replication process. Direct binding between these viral proteins was confirmed by mammalian cell two-hybrid analysis.

* Corresponding author. Mailing address: LID, NIAID, NIH, Bldg. 50, Room 6120, 50 South Drive, Bethesda, MD 20892-8011. Phone: (301) 594-1654. Fax: (301) 435-6021. E-mail: eehrenfeld{at}niaid.nih.gov.

Journal of Virology, June 2006, p. 5327-5337, Vol. 80, No. 11
0022-538X/06/$08.00+0     doi:10.1128/JVI.02684-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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