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J Virol, June 1998, p. 4783-4788, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Initiation of Hepatitis Delta Virus Genome Replication

Kate Dingle,1 Vadim Bichko,1 Harmon Zuccola,2 James Hogle,2 and John Taylor1,*

Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497,1 and Harvard Medical School, Boston, Massachusetts 021152

Received 11 December 1997/Accepted 3 March 1998

The small, 195-amino-acid form of the hepatitis delta virus (HDV) antigen (delta Ag-S) is essential for genome replication, i.e., for the transcription, processing, and accumulation of HDV RNAs. To better understand this requirement, we used purified recombinant delta Ag-S and HDV RNA synthesized in vitro to assemble high-molecular-weight ribonucleoprotein (RNP) structures. After transfection of these RNPs into human cells, we detected HDV genome replication, as assayed by Northern analysis or immunofluorescence microscopy. Our interpretation is that the input delta Ag-S is necessary for the RNA to undergo limited amounts of RNA-directed RNA synthesis, RNA processing, and mRNA formation, leading to de novo translation of delta Ag-S. It is this second source of delta Ag-S which then goes on to support genome replication. This assay made it possible to manipulate in vitro the composition of the RNP and then test in vivo the ability of the complex to initiate RNA-directed RNA synthesis and go on to achieve genome replication. For example, both genomic and antigenomic linear RNAs were acceptable. Substitution for delta Ag-S with truncated or modified forms of the delta Ag, and even with HIV nucleocapsid protein and polylysine, was unacceptable; the exception was a form of delta Ag-S with six histidines added at the C terminus. We expect that further in vitro modifications of these RNP complexes should help define the in vivo requirements for what we define as the initiation of HDV genome replication.

* Corresponding author. Mailing address: Fox Chase Cancer Center, 7701 Burholme Ave., Philadelphia, PA 19111-2497. Phone: (215) 728-2436. Fax: (215) 728-3616. E-mail: jm_taylor{at}fccc.edu.

J Virol, June 1998, p. 4783-4788, Vol. 72, No. 6
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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