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J Virol. 1990 November; 64(11): 5324–5332.
PMCID: PMC248581

The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation.

Abstract

Encapsidation of the pregenomic RNA into nucleocapsids is a selective process which depends on specific RNA-protein interactions. The signal involved in the packaging of the hepatitis B virus (HBV) RNA pregenome was recently defined as a short sequence located near the 5' end of that molecule (Junker-Niepmann et al., EMBO J., in press), but it remained an open question which viral proteins are required. Using a genetic approach, we analyzed whether proteins derived from the HBV P gene play an important role in pregenome encapsidation. The results obtained with point mutations, deletions, and insertions scattered throughout the P gene clearly demonstrate that (i) a P gene product containing all functional domains is required both for the encapsidation of HBV pregenomic RNA and for packaging of nonviral RNAs fused to the HBV encapsidation signal, (ii) known enzymatic activities are not involved in the packaging reaction, suggesting that P protein is required as a structural component, and (iii) P protein acts primarily in cis, i.e., pregenomic RNAs from which P protein is synthesized are preferentially encapsidated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bartenschlager R, Schaller H. The amino-terminal domain of the hepadnaviral P-gene encodes the terminal protein (genome-linked protein) believed to prime reverse transcription. EMBO J. 1988 Dec 20;7(13):4185–4192. [PubMed]
  • Cattaneo R, Will H, Hernandez N, Schaller H. Signals regulating hepatitis B surface antigen transcription. Nature. 1983 Sep 22;305(5932):336–338. [PubMed]
  • Challberg MD, Desiderio SV, Kelly TJ., Jr Adenovirus DNA replication in vitro: characterization of a protein covalently linked to nascent DNA strands. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5105–5109. [PubMed]
  • Chang LJ, Pryciak P, Ganem D, Varmus HE. Biosynthesis of the reverse transcriptase of hepatitis B viruses involves de novo translational initiation not ribosomal frameshifting. Nature. 1989 Jan 26;337(6205):364–368. [PubMed]
  • Galibert F, Mandart E, Fitoussi F, Tiollais P, Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. [PubMed]
  • Ganem D, Varmus HE. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. [PubMed]
  • Hirsch RC, Lavine JE, Chang LJ, Varmus HE, Ganem D. Polymerase gene products of hepatitis B viruses are required for genomic RNA packaging as wel as for reverse transcription. Nature. 1990 Apr 5;344(6266):552–555. [PubMed]
  • Jean-Jean O, Weimer T, de Recondo AM, Will H, Rossignol JM. Internal entry of ribosomes and ribosomal scanning involved in hepatitis B virus P gene expression. J Virol. 1989 Dec;63(12):5451–5454. [PMC free article] [PubMed]
  • Junker M, Galle P, Schaller H. Expression and replication of the hepatitis B virus genome under foreign promoter control. Nucleic Acids Res. 1987 Dec 23;15(24):10117–10132. [PMC free article] [PubMed]
  • Levin JG, Grimley PM, Ramseur JM, Berezesky IK. Deficiency of 60 to 70S RNA in murine leukemia virus particles assembled in cells treated with actinomycin D. J Virol. 1974 Jul;14(1):152–161. [PMC free article] [PubMed]
  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. [PMC free article] [PubMed]
  • Nassal M. Total chemical synthesis of a gene for hepatitis B virus core protein and its functional characterization. Gene. 1988 Jun 30;66(2):279–294. [PubMed]
  • Pasek M, Goto T, Gilbert W, Zink B, Schaller H, MacKay P, Leadbetter G, Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. [PubMed]
  • Radziwill G, Tucker W, Schaller H. Mutational analysis of the hepatitis B virus P gene product: domain structure and RNase H activity. J Virol. 1990 Feb;64(2):613–620. [PMC free article] [PubMed]
  • Roychoudhury S, Shih C. cis rescue of a mutated reverse transcriptase gene of human hepatitis B virus by creation of an internal ATG. J Virol. 1990 Mar;64(3):1063–1069. [PMC free article] [PubMed]
  • Schlicht HJ, Salfeld J, Schaller H. The duck hepatitis B virus pre-C region encodes a signal sequence which is essential for synthesis and secretion of processed core proteins but not for virus formation. J Virol. 1987 Dec;61(12):3701–3709. [PMC free article] [PubMed]
  • Schlicht HJ, Radziwill G, Schaller H. Synthesis and encapsidation of duck hepatitis B virus reverse transcriptase do not require formation of core-polymerase fusion proteins. Cell. 1989 Jan 13;56(1):85–92. [PubMed]
  • Schlicht HJ, Bartenschlager R, Schaller H. The duck hepatitis B virus core protein contains a highly phosphorylated C terminus that is essential for replication but not for RNA packaging. J Virol. 1989 Jul;63(7):2995–3000. [PMC free article] [PubMed]
  • Seeger C, Maragos J. Identification and characterization of the woodchuck hepatitis virus origin of DNA replication. J Virol. 1990 Jan;64(1):16–23. [PMC free article] [PubMed]
  • Shields A, Witte WN, Rothenberg E, Baltimore D. High frequency of aberrant expression of Moloney murine leukemia virus in clonal infections. Cell. 1978 Jul;14(3):601–609. [PubMed]
  • Sprengel R, Kuhn C, Will H, Schaller H. Comparative sequence analysis of duck and human hepatitis B virus genomes. J Med Virol. 1985 Apr;15(4):323–333. [PubMed]
  • Weimer T, Salfeld J, Will H. Expression of the hepatitis B virus core gene in vitro and in vivo. J Virol. 1987 Oct;61(10):3109–3113. [PMC free article] [PubMed]
  • Will H, Cattaneo R, Darai G, Deinhardt F, Schellekens H, Schaller H. Infectious hepatitis B virus from cloned DNA of known nucleotide sequence. Proc Natl Acad Sci U S A. 1985 Feb;82(3):891–895. [PubMed]
  • Will H, Salfeld J, Pfaff E, Manso C, Theilmann L, Schaler H. Putative reverse transcriptase intermediates of human hepatitis B virus in primary liver carcinomas. Science. 1986 Feb 7;231(4738):594–596. [PubMed]
  • Yaginuma K, Shirakata Y, Kobayashi M, Koike K. Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. Proc Natl Acad Sci U S A. 1987 May;84(9):2678–2682. [PubMed]
  • Zoller MJ, Smith M. Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template. DNA. 1984 Dec;3(6):479–488. [PubMed]

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