Organization and expression of the poxvirus genome

Wittek, Riccardo

doi:10.1007/bf01949349

Cited by 65 publications

(37 citation statements)

References 84 publications

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“…Because the HindIII restriction sites are well conserved among different vaccinia virus strains [36] our first step in characterizing the genome of the MVA virus was the comparison of its HindIII fi'agments with those of the W R virus D N A [7]. Southern blot analyses showed several differences between the DNAs ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Partial deletion of the human host range gene in the attenuated vaccinia virus MVA

Altenburger

Süter

Altenburger

1989

Archives of Virology

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The genome of a strongly attenuated vaccinia virus strain, MVA, was investigated by Southern blot and sequence analyses. Three major deletions, relative to the WR strain, were localized in MVA DNA. The deletions occurred near both ends of the viral genome and one of them affected a 55 K as well as the 32 K human host range gene. Although more than two thirds of the host range gene were eliminated from the MVA, the virus could still multiply in certain human cells.

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Section: Resultsmentioning

confidence: 99%

Partial deletion of the human host range gene in the attenuated vaccinia virus MVA

Altenburger

Süter

Altenburger

1989

Archives of Virology

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“…Late vaccinia virus transcripts are very heterogeneous in length (26), presumably as a consequence of incorrect termination of transcription late in infection. No attempt was made, therefore, to map the 3' end of the RNA.…”

Section: Resultsmentioning

confidence: 99%

One hundred base pairs of 5' flanking sequence of a vaccinia virus late gene are sufficient to temporally regulate late transcription.

Bertholet¹,

Drillien²,

Wittek³

1985

Proc. Natl. Acad. Sci. U.S.A.

114

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A vaccinia virus late gene coding for a major structural polypeptide of 11 kDa was sequenced. Although the 5' flanking gene region is very A+T rich, it shows little homology either to the corresponding region of vaccinia early genes or to consensus sequences characteristic of most eukaryotic genes. Three DNA fragments (100, 200, and 500 base pairs, respectively), derived from the flanking region and including the late gene mRNA start site, were inserted into the coding sequence of the vaccinia virus thymidine kinase (TK) early gene by homologous in vivo recombination. Recombinants were selected on the basis of their TK-phenotype. Cells were infected with the recombinant viruses and RNA was isolated at 1-hr intervals. Transcripts initiating either from the TK early promoter, or from the late gene promoter at its authentic position, or from the translocated late gene promoters within the early gene were detected by nuclease S1 mapping. Early after infection, only transcripts from the TK early promoter were detected. Later in infection, however, transcripts were also initiated from the translocated late promoters. This RNA appeared at the same time and in similar quantities as the RNA from the late promoter at its authentic position. No quantitative differences in promoter efficiency between the 100-, 200-, and 500-base-pair insertions were observed. We conclude that all necessary signals for correct regulation of late-gene expression reside within only 100 base pairs of 5' flanking sequence.Vaccinia virus, a member of the poxvirus family, contains a large double-stranded DNA genome of 180 kilobase pairs. Expression of this large amount of genetic information is temporally well-regulated. Early genes are transcribed shortly after penetration of the virus particles into the host cell. After DNA replication, late genes encoding predominantly structural polypeptides are expressed. The molecular basis for this temporal regulation is not understood.In contrast to other animal DNA viruses, which replicate in the nucleus of the infected cells and which use the host cell RNA polymerase to transcribe their genes, vaccinia replicates in the cytoplasm and utilizes its own transcription system. A multisubunit RNA polymerase (1, 2) and enzymes involved in modification of RNA (3, 4) have been isolated from purified virus particles. The mRNAs made by these enzymes are not spliced (5, 6) but have cap structures (7) and poly(A) tails (8), which are characteristic features of eukaryotic mRNAs. As might be expected, recent evidence suggests that vaccinia virus has evolved its own regulatory signals for gene expression, First, vaccinia genes are transcribed in cell-free extracts prepared from infected cells but not in extracts from non-infected cells (9), which suggests that the host-cell transcriptional machinery does not recognize vaccinia promoter elements. Second, the 5' flanking regions of four early genes that have been sequenced (10-13) lack the regulatory elements characteristic of the corresponding regions of most cel...

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“…Orthopoxviruses have .been standardized mainly by biological characteristics and DNA analysis. Quantitative serological differentiation by polyclonal antibodies is difficult and, because of their close relationships, not very helpful (Mahnel et al, 1974;Wittek, 1982;Esposito & Knight, 1985). ELISAs are well suited to evaluating qualitative and quantitative epitope characteristics in virus strains.…”

Section: Discussionmentioning

confidence: 99%

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Czerny¹,

Mahnel²

1990

Journal of General Virology

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Organization and expression of the poxvirus genome

Cited by 65 publications

References 84 publications

Partial deletion of the human host range gene in the attenuated vaccinia virus MVA

Partial deletion of the human host range gene in the attenuated vaccinia virus MVA

One hundred base pairs of 5' flanking sequence of a vaccinia virus late gene are sufficient to temporally regulate late transcription.

Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

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