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Journal of Virology, December 1998, p. 9668-9675, Vol. 72, No. 12
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.
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
and
*
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.
Present address: ISIS Pharmaceuticals, Carlsbad, CA 92008.
Present address: National Institutes of Health, National Institute
of Allergy and Infectious Diseases, LVD, Picornavirus Section, Bethesda, MD 20892.
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