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Function of rotavirus VP3 polypeptide in viral morphogenesis
M Vásquez1, A M Sandino, J M Pizarro
1Instituto de Nutrición y Tecnología de los Alimentos (INTA), Universidad de Chile, Santiago.
The Journal of General Virology
|May 1, 1993
Summary
Rotavirus SA-11 mutant tsB, with a thermosensitive mutation in gene 3 (VP3), halts morphogenesis before RNA replication. This suggests VP3 is crucial for RNA replication, not viral assembly.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Rotavirus SA-11 mutant tsB possesses a thermosensitive mutation in gene 3, encoding viral protein 3 (VP3).
- The mutant's phenotype was initially characterized as lacking in vivo RNA synthesis.
Purpose of the Study:
- To further characterize the phenotype of the rotavirus SA-11 mutant tsB.
- To investigate the role of VP3 in rotavirus RNA synthesis and viral morphogenesis.
Main Methods:
- Characterization of infected cells and purified viral particles.
- In vitro transcriptional assays at restrictive temperatures.
- Analysis of VP3-associated guanylyl-transferase activity.
- Morphological analysis of assembled viral particles.
Main Results:
- The tsB mutant's in vitro transcription and VP3 guanylyl-transferase activity were identical to the wild-type strain at restrictive temperatures.
- Viral particles assembled by mutant-infected cells at restrictive temperatures were empty, single-shelled particles.
- Viral morphogenesis halts after transcription but before RNA replication.
Conclusions:
- The observed phenotype is not due to altered VP3 function in transcription.
- VP3 is likely essential for the RNA replication machinery.
- VP3 is not required for the assembly of subviral particles.