Identification of the functional regions required for hepatitis D virus replication and transcription by linker-scanning mutagenesis of viral genome

Virology
H W WangP J Chen

Abstract

To define the important cis-elements in hepatitis delta virus (HDV) RNA, the viral genome was mutated by a linker-scanning mutagenesis strategy that maintained the native rod-like structure of HDV RNA. Mutant HDV cDNAs or their corresponding RNA transcripts were transfected into a Huh-7-derived cell line which continuously expressed small hepatitis delta antigen to study the viral replication and transcription. Here we report the following findings. (i) Although most of the mutant RNAs could self-process to generate the 1.7-kb genomic RNA and all their stabilities were similar, positions which surround the genomic ribozyme domain were found to be important for the self-processing of the dimeric RNA. (ii) The replication of viral RNA was greatly diminished in many mutants, suggesting that multiple regions in HDV RNA were required for replication. (iii) In certain mutants, replication of the HDV antigenomic RNA was selectively abolished but that of the genomic RNA was not. Therefore, this was the first report to show that the cis-elements needed for the replication of genomic or antigenomic HDV RNA could be different. (iv) A continuous region (nt 1625 to nt 431), spanning the HDAg mRNA initiation site and containing the in vitro ...Continue Reading

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Citations

Feb 11, 2000·RNA·J Filipovska, M M Konarska
Sep 10, 2005·Nucleic Acids Research·Subash C B GopinathPenmetcha K R Kumar
Jul 25, 2000·Journal of Virology·S GudimaJ Taylor
Apr 19, 2006·RNA·Severin O GudimaJohn M Taylor
Mar 10, 2010·Future Microbiology·John Taylor, Martin Pelchat
Sep 19, 2007·Proceedings of the National Academy of Sciences of the United States of America·Philip Downey
Sep 24, 2004·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Shiou-Hwei YehPei-Jer Chen
Oct 1, 2010·Liver International : Official Journal of the International Association for the Study of the Liver·Stéphanie Pascarella, Francesco Negro
Mar 29, 2000·Critical Reviews in Clinical Laboratory Sciences·L E Modahl, M M Lai
Oct 1, 2010·Molecular Cancer Research : MCR·Subash C B GopinathPenmetcha K R Kumar

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