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Proteins expressed by Mill Door/79 virus, a Kemerovo serogroup orbivirus transmitted by the ticks Ixodes uriae
Abstract:
Mill Door/79 virus, an orbivirus of the Kemerovo serogroup, Great Island Complex, was shown to induce in infected cells 10 segments of double-stranded RNA with a total molecular weight of 11.46 X 10(6) daltons. Using methyl mercuric hydroxide as a denaturing agent the double-stranded RNA was translated in a cell-free system producing 11 polypeptides, 10 of which co-migrated with those produced in Mill Door/79 virus infected Vero cells. The segments were separated and individually translated in a cell-free system allowing their coding assignments to be made. These assignments were confirmed by partial proteolysis.
Insights
Mill Door/79 virus infection produces 10 double-stranded RNA segments. Cell-free translation identified 11 polypeptides, mapping viral gene coding assignments confirmed by proteolysis.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Orbiviruses, including the Kemerovo serogroup, are significant viral pathogens.
- Understanding the genome and protein expression of novel orbiviruses is crucial for disease control.
- Mill Door/79 virus is a newly identified orbivirus belonging to the Great Island Complex.
Purpose of the Study:
- To characterize the double-stranded RNA genome of Mill Door/79 virus.
- To identify and map the viral proteins encoded by the Mill Door/79 virus genome.
- To elucidate the genetic basis of Mill Door/79 virus replication and pathogenesis.
Main Methods:
- Infection of Vero cells with Mill Door/79 virus.
- Extraction and denaturation of viral double-stranded RNA using methyl mercuric hydroxide.
- Cell-free translation of dsRNA segments.
- Separation and individual translation of dsRNA segments.
- Analysis of translated polypeptides by co-migration and partial proteolysis.
Main Results:
- Mill Door/79 virus infection induced 10 double-stranded RNA segments with a total molecular weight of 11.46 x 10(6) daltons.
- Cell-free translation of denatured dsRNA yielded 11 polypeptides.
- Ten of these polypeptides co-migrated with proteins produced in infected Vero cells.
- Individual translation of separated dsRNA segments allowed for coding assignments.
- Partial proteolysis confirmed the coding assignments of viral gene products.
Conclusions:
- The double-stranded RNA genome of Mill Door/79 virus encodes at least 11 distinct viral proteins.
- The study successfully mapped the coding potential of the Mill Door/79 virus genome.
- This provides a foundation for further research into orbivirus replication and the development of diagnostic and therapeutic strategies.