This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gudima, S. Articles by Taylor, J. Search for Related Content PubMed PubMed Citation Articles by Gudima, S. Articles by Taylor, J.

 Previous Article  |  Next Article 

Journal of Virology, April 2002, p. 3709-3719, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.3709-3719.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Parameters of Human Hepatitis Delta Virus Genome Replication: the Quantity, Quality, and Intracellular Distribution of Viral Proteins and RNA

Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497

Received 2 November 2001/ Accepted 2 January 2002

Assembly of hepatitis delta virus (HDV) in infected human hepatocytes involves association of the 1,679- nucleotide single-stranded genomic RNA (RNA) with multiple copies of both small and large forms of the delta protein (Ag) to form a ribonucleoprotein particle which in turn interacts with envelope proteins of the natural helper virus, hepatitis B virus. Subsequently, for initiation of a new round of replication, the amount of small Ag within the assembled HDV particle is both necessary and sufficient. Quantitative assays were used in order to better understand just how much Ag is needed. The molar ratio of Ag species to genomic RNA in assembled HDV particles was approximately 200. Next, this ratio was determined for cells under several different experimental situations in which HDV genome replication was occurring. These included replication in woodchuck liver and also in mouse liver and skeletal muscle, as well as replication in stably and transiently transfected cultured human hepatoblastoma cells. Surprisingly, in almost all these situations the molar ratios were comparable to that observed for HDV particles. This was true for different times after the initiation of replication and was independent of whether or not virus assembly was occurring. Cell fractionation combined with quantitative assays was used to test whether the majority of Ag and RNA were colocalized during HDV replication in transfected cells. The cytoplasmic fraction contained the majority of Ag and genomic RNA. Finally, the quality of Ag and RNA, especially at relatively late times after the initiation of replication, was examined by using reverse transcription-PCR, cloning, and sequencing through the entire Ag open reading frame. When virus assembly and spread were not possible, 20% or less of the predicted Ag would have been able to support HDV replication. In summary, an examination of the quantity, quality and intracellular distribution of Ag and RNA in several different experimental systems has provided a better understanding of the parameters associated with the initiation, maintenance, and ultimate decline of HDV genome replication.

This article has been cited by other articles:

• Chang, J., Nie, X., Chang, H. E., Han, Z., Taylor, J. (2008). Transcription of Hepatitis Delta Virus RNA by RNA Polymerase II. J. Virol. 82: 1118-1127 [Abstract] [Full Text]  
• Gudima, S., He, Y., Meier, A., Chang, J., Chen, R., Jarnik, M., Nicolas, E., Bruss, V., Taylor, J. (2007). Assembly of Hepatitis Delta Virus: Particle Characterization, Including the Ability To Infect Primary Human Hepatocytes. J. Virol. 81: 3608-3617 [Abstract] [Full Text]  
• Gudima, S. O., Chang, J., Taylor, J. M. (2006). Restoration in vivo of defective hepatitis delta virus RNA genomes. RNA 12: 1061-1073 [Abstract] [Full Text]  
• Chang, J., Gudima, S. O., Taylor, J. M. (2005). Evolution of Hepatitis Delta Virus RNA Genome following Long-Term Replication in Cell Culture. J. Virol. 79: 13310-13316 [Abstract] [Full Text]  
• Chang, J., Gudima, S. O., Tarn, C., Nie, X., Taylor, J. M. (2005). Development of a Novel System To Study Hepatitis Delta Virus Genome Replication. J. Virol. 79: 8182-8188 [Abstract] [Full Text]  
• GUDIMA, S. O., CHANG, J., TAYLOR, J. M. (2005). Reconstitution in cultured cells of replicating HDV RNA from pairs of less than full-length RNAs. RNA 11: 90-98 [Abstract] [Full Text]  
• Gudima, S. O., Chang, J., Taylor, J. M. (2004). Features Affecting the Ability of Hepatitis Delta Virus RNAs To Initiate RNA-Directed RNA Synthesis. J. Virol. 78: 5737-5744 [Abstract] [Full Text]  
• Cornillez-Ty, C. T., Lazinski, D. W. (2003). Determination of the Multimerization State of the Hepatitis Delta Virus Antigens In Vivo. J. Virol. 77: 10314-10326 [Abstract] [Full Text]  
• Mankertz, A., Mueller, B., Steinfeldt, T., Schmitt, C., Finsterbusch, T. (2003). New Reporter Gene-Based Replication Assay Reveals Exchangeability of Replication Factors of Porcine Circovirus Types 1 and 2. J. Virol. 77: 9885-9893 [Abstract] [Full Text]  
• Chang, J., Taylor, J. M. (2003). Susceptibility of Human Hepatitis Delta Virus RNAs to Small Interfering RNA Action. J. Virol. 77: 9728-9731 [Abstract] [Full Text]  
• Wong, S. K., Lazinski, D. W. (2002). Replicating hepatitis delta virus RNA is edited in the nucleus by the small form of ADAR1. Proc. Natl. Acad. Sci. USA 99: 15118-15123 [Abstract] [Full Text]