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Journal of Virology, December 2002, p. 12457-12462, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12457-12462.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Cytoplasmic Expression of mRNAs Containing the Internal Ribosome Entry Site and 3' Noncoding Region of Hepatitis C Virus: Effects of the 3' Leader on mRNA Translation and mRNA Stability

Li Kuo Kong1,2,3 and Peter Sarnow1*

Departments of Microbiology and Immunology,1 Medicine,2 Infectious Diseases, Stanford University School of Medicine, Stanford, California 943053

Received 9 July 2002/ Accepted 12 September 2002

Translation initiation in many eukaryotic mRNAs is modulated by an interaction between the cap binding protein complex, bound to the 5' end of the mRNA, and the polyadenosine binding protein, bound to the 3'-terminal polyadenosine sequences. A few cellular and viral mRNAs, such as the hepatitis C virus (HCV) mRNA genome, lack 3'-terminal polyadenosine sequences. For such mRNAs, the question of whether their 3'-end sequences also regulate the initiation phase of protein synthesis via an interaction with their 5' ends has received intense scrutiny. For HCV mRNA, various experimental designs have led to conflicting interpretations, that the 3' end of the RNA can modulate translation initiation either in a positive or in a negative fashion. To examine the possibility of end-to-end communication in HCV in detail, mRNAs containing the HCV internal ribosome entry site linked to a luciferase coding region, followed by different 3' noncoding regions, were expressed in the cytoplasm of cultured cells by T7 RNA polymerase. The intracellular translation efficiencies, steady-state levels, stabilities, and 3'-end sequences of these chimeric RNAs were examined. It was found that the HCV 3' noncoding region modulates neither the translation nor the stability of the mRNAs. Thus, there is no detectable end-to-end communication in cytoplasmically expressed chimeric mRNAs containing the HCV noncoding regions. However, it remains an open question whether end-to-end communication occurs in full-length HCV mRNAs in the infected liver.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Stanford University School of Medicine, Sherman Fairchild Science Bldg., 299 Campus Dr., Stanford, CA 94305-5124. Phone: (650) 498-7076. Fax: (650) 498-7147. E-mail: psarnow{at}leland.stanford.edu.

Journal of Virology, December 2002, p. 12457-12462, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12457-12462.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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