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Rotavirus RNA replication: single-stranded RNA extends from the replicase particle.

J T Patton¹, C O Gallegos

¹Department of Microbiology and Immunology, University of Miami School of Medicine, Florida 33101.

The Journal of General Virology

|May 1, 1990

Summary

Rotavirus replicase particles change size during RNA replication, decreasing in size as more double-stranded RNA (dsRNA) is synthesized. This structural change is linked to the template RNA extending from the particle.

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Area of Science:

Virology
Molecular Biology
RNA Replication

Background:

Rotavirus possesses an 11-segment double-stranded RNA (dsRNA) genome.
dsRNA replication occurs within subviral particles (SVPs).
The precise structural dynamics of replicase particles during RNA synthesis remain incompletely understood.

Purpose of the Study:

To investigate structural changes in rotavirus replicase particles during dsRNA synthesis.
To elucidate the relationship between replicase particle size and RNA replication activity.

Main Methods:

Non-denaturing gel electrophoresis of purified SVPs from infected cells.
Cell-free system for dsRNA synthesis by replicase particles.
Analysis of particle size over time during incubation.

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RNase treatment to assess template RNA accessibility.

Main Results:

Replicase particles synthesizing dsRNA were initially large (≥100 nm).
Particle size decreased with extended incubation, correlating with increased dsRNA production.
Final particle size resembled previously described rotavirus replicative intermediates.
RNase sensitivity indicated the plus-strand RNA template extends externally.

Conclusions:

Rotavirus replicase particles undergo dynamic size changes during dsRNA replication.
This size modulation is driven by the incorporation of plus-strand RNA templates.
Understanding these dynamics offers insights into rotavirus RNA synthesis regulation.