Abstract
Unlike many other reverse transcriptase genes, the polymerase (P) gene of the hepatitis B viruses is expressed by translational initiation from its own AUG codon rather than by ribosomal frameshifting during translation of the overlapping core gene (C). To explore the mechanism of its translation, we have fused the P gene of duck hepatitis B virus to the bacterial lacZ gene at a point 3' to the C-P overlap; this allows detection of the products of P translation with antisera to the lacZ-encoded protein. The C and P/Z coding regions were cloned downstream of a heterologous promoter and expressed in COS-7 cells. A single, bicistronic mRNA containing both C and P sequences is detected in these cells, and translational initiation occurs efficiently at the internally situated P AUG. Mutations affecting C translation have only minimal effects on P expression, in contrast to what would be expected from a modified scanning model for translation. We conclude that P translation depends on a mechanism other than scanning to allow internal entry of ribosomes to the region of the P initiator.
MeSH Terms
Animals
Base Sequence
Cell Line
Codon/genetics
Genes, Viral
Hepadnaviridae/enzymology,genetics
Molecular Sequence Data
Mutation
Plasmids
Poliovirus/genetics
Protein Biosynthesis
RNA, Messenger/genetics
RNA-Directed DNA Polymerase/genetics,metabolism
Recombinant Fusion Proteins/metabolism
Viral Structural Proteins/genetics
Chemicals
Codon
RNA, Messenger
Recombinant Fusion Proteins
Viral Structural Proteins
RNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Chang L J
Department of Microbiology and Immunology, University of California, San Francisco 94143.
Ganem D
Varmus H E
References (18)
18 references, click to expand
-
Novel N-terminal amino acid sequence required for retention of a hepatitis B virus glycoprotein in the endoplasmic reticulum.
Mol Cell Biol. 1989 Oct;9(10):4459-66
PMID: 2586518
-
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
PMID: 2463093
-
Ty: a retroelement moving forward.
Cell. 1988 May 6;53(3):333-5
PMID: 2452694
-
Transcripts and the putative RNA pregenome of duck hepatitis B virus: implications for reverse transcription.
Cell. 1985 Mar;40(3):717-24
PMID: 2857595
-
Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA.
Nature. 1988 Jul 28;334(6180):320-5
PMID: 2839775
-
Regulation of protein synthesis in virus-infected animal cells.
Adv Virus Res. 1986;31:229-92
PMID: 3019107
-
A cis-acting element within the 5' leader of a cytomegalovirus beta transcript determines kinetic class.
Cell. 1986 Sep 12;46(6):865-72
PMID: 3019554
-
Biosynthesis of the reverse transcriptase of hepatitis B viruses involves de novo translational initiation not ribosomal frameshifting.
Nature. 1989 Jan 26;337(6205):364-8
PMID: 2463489
-
Expression of the Rous sarcoma virus pol gene by ribosomal frameshifting.
Science. 1985 Dec 13;230(4731):1237-42
PMID: 2416054
-
The scanning model for translation: an update.
J Cell Biol. 1989 Feb;108(2):229-41
PMID: 2645293
-
Internal entry of ribosomes and ribosomal scanning involved in hepatitis B virus P gene expression.
J Virol. 1989 Dec;63(12):5451-4
PMID: 2585611
-
Comparative sequence analysis of duck and human hepatitis B virus genomes.
J Med Virol. 1985 Apr;15(4):323-33
PMID: 3981148
-
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):552-5
PMID: 1690862
-
Construction of a modular dihydrofolate reductase cDNA gene: analysis of signals utilized for efficient expression.
Mol Cell Biol. 1982 Nov;2(11):1304-19
PMID: 6131378
-
SV40-transformed simian cells support the replication of early SV40 mutants.
Cell. 1981 Jan;23(1):175-82
PMID: 6260373
-
Initiation of translation at internal AUG codons in mammalian cells.
Nature. 1984 May 3-9;309(5963):82-5
PMID: 6717585
-
Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences.
J Virol. 1984 Mar;49(3):782-92
PMID: 6699938
-
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
Science. 1988 Jan 29;239(4839):487-91
PMID: 2448875