Complementation Analysis of Mutants Defective in Different Steps of HBV Reverse Transcription

Overview

Journal Gastroenterol Jpn

Specialty Gastroenterology

Date 1990 Sep 1

PMID 2227268

Authors

H Schaller

G Radziwill

Affiliations

Soon will be listed here.

Abstract

The HBV P gene encodes a multifunctional polyprotein which contains the major enzymatic activities required for hepadnaviral reverse transcription (protein primer, DNA polymerase, and RNase H). Mutant HBV genomes unable to synthesize fully functional P gene products were analysed for their potential to be rescued by a second mutant HBV genome that either contained a wild type P gene (intergenic complementation) or a mutation in a functionally different P gene domain (intragenic complementation). This analysis was carried out by cotransfecting two mutants at a time into HepG2 cells and assaying for the production of core particles containing DNA polymerase activity. The results obtained demonstrate the existence of intergenic, but not of intragenic complementation. This indicates that the primary P gene product is not processed before core assembly, and furthermore that there is a rather free mixing of all HBV gene products in the HBV infected cell which can lead to the efficient propagation of defective viral genomes.

References

Jacks T, Varmus H . Expression of the Rous sarcoma virus pol gene by ribosomal frameshifting. Science. 1985; 230(4731):1237-42. DOI: 10.1126/science.2416054. View

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; 231(4738):594-6. DOI: 10.1126/science.2418501. View

Schlicht H, 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; 56(1):85-92. DOI: 10.1016/0092-8674(89)90986-0. View

Chang L, Pryciak P, Ganem D, Varmus H . Biosynthesis of the reverse transcriptase of hepatitis B viruses involves de novo translational initiation not ribosomal frameshifting. Nature. 1989; 337(6205):364-8. DOI: 10.1038/337364a0. View

Galle P, Schlicht H, Fischer M, Schaller H . Production of infectious duck hepatitis B virus in a human hepatoma cell line. J Virol. 1988; 62(5):1736-40. PMC: 253216. DOI: 10.1128/JVI.62.5.1736-1740.1988. View

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; 7(13):4185-92. PMC: 455131. DOI: 10.1002/j.1460-2075.1988.tb03315.x. View

Acs G, Sells M, Purcell R, Price P, Engle R, Shapiro M . Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees. Proc Natl Acad Sci U S A. 1987; 84(13):4641-4. PMC: 305146. DOI: 10.1073/pnas.84.13.4641. View

Sureau C, Mullins J, Essex M . Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell. 1986; 47(1):37-47. DOI: 10.1016/0092-8674(86)90364-8. View

Tsurimoto T, Fujiyama A, Matsubara K . Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc Natl Acad Sci U S A. 1987; 84(2):444-8. PMC: 304224. DOI: 10.1073/pnas.84.2.444. View

10.

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; 84(9):2678-82. PMC: 304721. DOI: 10.1073/pnas.84.9.2678. View

11.

Chang C, Jeng K, Hu C, Lo S, Su T, Ting L . Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. EMBO J. 1987; 6(3):675-80. PMC: 553450. DOI: 10.1002/j.1460-2075.1987.tb04807.x. View

12.

Popper H, Shafritz D, Hoofnagle J . Relation of the hepatitis B virus carrier state to hepatocellular carcinoma. Hepatology. 1987; 7(4):764-72. DOI: 10.1002/hep.1840070425. View

13.

Ganem D, Varmus H . The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987; 56:651-93. DOI: 10.1146/annurev.bi.56.070187.003251. View

14.

Okamoto H, Tsuda F, Mayumi M . Defective mutants of hepatitis B virus in the circulation of symptom-free carriers. Jpn J Exp Med. 1987; 57(4):217-21. View

15.

Schlicht H, 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; 61(12):3701-9. PMC: 255982. DOI: 10.1128/JVI.61.12.3701-3709.1987. View

16.

Graham F, Van Der Eb A . A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973; 52(2):456-67. DOI: 10.1016/0042-6822(73)90341-3. View

17.

Summers J, Mason W . Replication of the genome of a hepatitis B--like virus by reverse transcription of an RNA intermediate. Cell. 1982; 29(2):403-15. DOI: 10.1016/0092-8674(82)90157-x. View

18.

Toh H, Hayashida H, Miyata T . Sequence homology between retroviral reverse transcriptase and putative polymerases of hepatitis B virus and cauliflower mosaic virus. Nature. 1983; 305(5937):827-9. DOI: 10.1038/305827a0. View

19.

Challberg M, Desiderio S, Kelly Jr T . Adenovirus DNA replication in vitro: characterization of a protein covalently linked to nascent DNA strands. Proc Natl Acad Sci U S A. 1980; 77(9):5105-9. PMC: 350005. DOI: 10.1073/pnas.77.9.5105. View