Detection of RNA-protein complex in vaccinia virus core in vitro transcription system

Damaso, Clarissa R.; Moussatché, Nissin

doi:10.1099/0022-1317-73-5-1243

Cited by 3 publications

(1 citation statement)

References 36 publications

(32 reference statements)

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“…At 1, 3, 6 and 8 h post‐treatment, the monolayers were harvested in TENS buffer (100 m m Tris–HCl, pH 8·0; 1 m m ethylenediaminetetraacetic acid, pH 8·0; 100 m m NaCl and 1% SDS), and equal volume of 30% cold trichloroacetic acid was added for 30 min at 4°C. The precipitate was collected on a glass‐fibre filter (GF/C; Whatman, Clifton, NJ, USA), and then the radioactivity present on the filters was determined by scintillation counting (Damaso and Moussatche 1992b).…”

Section: Rna Synthesis Analysismentioning

confidence: 99%

Protein synthesis inhibitory activity in culture filtrates from new strains of Streptomyces isolated from Brazilian tropical soils

Reis

Costa

Cavalcanti

et al. 2003

Lett Appl Microbiol

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Aims:To investigate the effect of the culture supernatants from three newly isolated Streptomyces strains, 221, 235 and 606 on eukaryotic cells. Methods and Results: Cell lines were treated with the culture filtrates and assayed for protein synthesis by metabolic labelling, followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis. RNA synthesis was investigated by [5-3 H]uridine incorporation. The three culture filtrates presented a strong inhibitory activity, reducing total protein synthesis of different eukaryotic cell lines by more than 85%. No effect on cellular RNA synthesis was detected. The culture filtrates did not affect the growth of the prokaryotic cells tested. Conclusions: These new Streptomyces strains, recently isolated from Brazilian tropical soils, produce molecule(s) with inhibitory activity specific to eukaryote protein synthesis. Significance and Impact of the Study: Streptomyces strains 221, 235 and 606, probably representing new species, might produce new bioactive compound(s), and can be used as valuable tools to study the protein synthesis pathway in eukaryotes.

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Section: Rna Synthesis Analysismentioning

confidence: 99%

Protein synthesis inhibitory activity in culture filtrates from new strains of Streptomyces isolated from Brazilian tropical soils

Reis

Costa

Cavalcanti

et al. 2003

Lett Appl Microbiol

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Azathioprine Inhibits Vaccinia Virus Replication in Both BSC-40 and Rag Cell Lines Acting on Different Stages of Virus Cycle

et al. 2002

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In the present study we demonstrate that azathioprine (AZA) inhibits vaccinia virus (VV) replication in both BSC-40 and RAG cell lines, acting on different stages of virus cycle. In BSC-40 cells, early protein synthesis was not significantly affected, but late gene expression was severely impaired. In RAG cells all stages of gene expression were completed during synchronous infection in the presence of the drug. The onset of DNA replication was not affected in RAG cells, but a severe inhibition was observed in BSC-40 cells. Electron microscopic analysis of VV-infected RAG cells treated with AZA revealed brick-shaped particles presenting abnormal definition of the internal structure. Purified virions from AZA-treated RAG cells presented several modifications of the protein content, a lesser amount of DNA, and a lower PFU:particle ratio. Our results suggest that in VV-infected RAG cells AZA interfered with virus morphogenesis, whereas in BSC-40 cells the replicative cycle was inhibited at the DNA replication stage.

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Cidofovir Inhibits Genome Encapsidation and Affects Morphogenesis during the Replication of Vaccinia Virus

Jesus

Costa

Gonçalves

et al. 2009

J Virol

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Cidofovir (CDV) is one of the most effective antiorthopoxvirus drugs, and it is widely accepted that viral DNA replication is the main target of its activity. In the present study, we report a detailed analysis of CDV effects on the replicative cycles of distinct vaccinia virus (VACV) strains: Cantagalo virus, VACV-IOC, and VACV-WR. We show that despite the approximately 90% inhibition of production of virus progeny, virus DNA accumulation was reduced only 30%, and late gene expression and genome resolution were unaltered. The level of proteolytic cleavage of the major core proteins was diminished in CDV-treated cells. Electron microscopic analysis of virus-infected cells in the presence of CDV revealed reductions as great as 3.5-fold in the number of mature forms of virus particles, along with a 3.2-fold increase in the number of spherical immature particles. A detailed analysis of purified virions recovered from CDV-treated cells demonstrated the accumulation of unprocessed p4a and p4b and nearly 67% inhibition of DNA encapsidation. However, these effects of CDV on virus morphogenesis resulted from a primary effect on virus DNA synthesis, which led to later defects in genome encapsidation and virus assembly. Analysis of virus DNA by atomic force microscopy revealed that viral cytoplasmic DNA synthesized in the presence of CDV had an altered structure, forming aggregates with increased strand overlapping not observed in the absence of the drug. These aberrant DNA aggregations were not encapsidated into virus particles.

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Detection of RNA-protein complex in vaccinia virus core in vitro transcription system

Cited by 3 publications

References 36 publications

Protein synthesis inhibitory activity in culture filtrates from new strains of Streptomyces isolated from Brazilian tropical soils

Protein synthesis inhibitory activity in culture filtrates from new strains of Streptomyces isolated from Brazilian tropical soils

Azathioprine Inhibits Vaccinia Virus Replication in Both BSC-40 and Rag Cell Lines Acting on Different Stages of Virus Cycle

Cidofovir Inhibits Genome Encapsidation and Affects Morphogenesis during the Replication of Vaccinia Virus

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