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Journal of Virology, February 2004, p. 1344-1351, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1344-1351.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cell-Dependent Role for the Poliovirus 3' Noncoding Region in Positive-Strand RNA Synthesis

David M. Brown,1,{dagger} Steven E. Kauder,2 Christopher T. Cornell,1,{ddagger} Gwendolyn M. Jang,1 Vincent R. Racaniello,2 and Bert L. Semler1*

Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine, California 92697,1 Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 008582

Received 22 May 2003/ Accepted 23 October 2003

We previously reported the isolation of a mutant poliovirus lacking the entire genomic RNA 3' noncoding region. Infection of HeLa cell monolayers with this deletion mutant revealed only a minor defect in the levels of viral RNA replication. To further analyze the consequences of the genomic 3' noncoding region deletion, we examined viral RNA replication in a neuroblastoma cell line, SK-N-SH cells. The minor genomic RNA replication defect in HeLa cells was significantly exacerbated in the SK-N-SH cells, resulting in a decreased capacity for mutant virus growth. Analysis of the nature of the RNA replication deficiency revealed that deleting the poliovirus genomic 3' noncoding region resulted in a positive-strand RNA synthesis defect. The RNA replication deficiency in SK-N-SH cells was not due to a major defect in viral translation or viral protein processing. Neurovirulence of the mutant virus was determined in a transgenic mouse line expressing the human poliovirus receptor. Greater than 1,000 times more mutant virus was required to paralyze 50% of inoculated mice, compared to that with wild-type virus. These data suggest that, together with a cellular factor(s) that is limiting in neuronal cells, the poliovirus 3' noncoding region is involved in positive-strand synthesis during genome replication.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, College of Medicine, Med. Sci. B240, University of California, Irvine, CA 92697-4025. Phone: (949) 824-7573. Fax: (949) 824-2694. E-mail: blsemler{at}uci.edu.

{dagger} Present address: Department of Molecular Biology, Princeton University, Princeton, NJ 08544.

{ddagger} Present address: Department of Neuropharmacology, Scripps Research Institute, La Jolla, CA 92037.

Journal of Virology, February 2004, p. 1344-1351, Vol. 78, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.3.1344-1351.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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