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Journal of Virology, December 1998, p. 9668-9675, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Interaction of Poly(rC) Binding Protein 2 with the 5' Noncoding Region of Hepatitis A Virus RNA and Its Effects on Translation

Judith Graff,* John Cha, Lawrence B. Blyn,dagger and Ellie EhrenfeldDagger

Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697

Received 26 March 1998/Accepted 24 August 1998

Utilization of internal ribosome entry segment (IRES) structures in the 5' noncoding region (5'NCR) of picornavirus RNAs for initiation of translation requires a number of host cell factors whose distribution may vary in different cells and whose requirement may vary for different picornaviruses. We have examined the requirement of the cellular protein poly(rC) binding protein 2 (PCBP2) for hepatitis A virus (HAV) RNA translation. PCBP2 has recently been identified as a factor required for translation and replication of poliovirus (PV) RNA. PCBP2 was shown to be present in FRhK-4 cells, which are permissive for growth of HAV, as it is in HeLa cells, which support translation of HAV RNA but which have not been reported to host replication of the virus. Competition RNA mobility shift assays showed that the 5'NCR of HAV RNA competed for binding of PCBP2 with a probe representing stem-loop IV of the PV 5'NCR. The binding site on HAV RNA was mapped to nucleotides 1 to 157, which includes a pyrimidine-rich sequence. HeLa cell extracts that had been depleted of PCBP2 by passage over a PV stem-loop IV RNA affinity column supported only low levels of HAV RNA translation. Translation activity was restored upon addition of recombinant PCBP2 to the depleted extract. Removal of the 5'-terminal 138 nucleotides of the HAV RNA, or removal of the entire IRES, eliminated the dependence of HAV RNA translation on PCBP2.

* Corresponding author. Mailing address: National Institutes of Health, NIAID, LID, Molecular Hepatitis Section, Building 7, Room 200, 7 Center Dr., Bethesda, MD 20892-0740. Phone: (301) 496-6227. Fax: (301) 402-0524. E-mail: jgraff{at}atlas.niaid.nih.gov.

dagger Present address: ISIS Pharmaceuticals, Carlsbad, CA 92008.

Dagger Present address: National Institutes of Health, National Institute of Allergy and Infectious Diseases, LVD, Picornavirus Section, Bethesda, MD 20892.

Journal of Virology, December 1998, p. 9668-9675, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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