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Characterization of rubella virus replication complexes using antibodies to double-stranded RNA
J Y Lee1, J A Marshall, D S Bowden
1Macfarlane Burnet Centre for Medical Research, Fairfield Hospital, Victoria, Australia.
Virology
|April 1, 1994
Summary
Rubella virus (RV) and Semliki Forest virus (SFV) replication complexes contain double-stranded RNA (dsRNA) within vesicles. These vesicles release amorphous material containing dsRNA during the viral replication cycle.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Rubella virus (RV) replication involves cytoplasmic structures called replication complexes.
- These complexes are crucial for viral RNA synthesis and assembly.
Purpose of the Study:
- To characterize the rubella virus replication complex.
- To identify the location and nature of double-stranded RNA (dsRNA) within these complexes.
Main Methods:
- Pre-embedding immunogold labeling electron microscopy was used on detergent-treated RV-infected cells.
- Antiserum to double-stranded RNA (dsRNA) was employed for labeling.
- Cells infected with Semliki Forest virus (SFV) were also examined for comparison.
Main Results:
- Gold particles, indicating dsRNA, were concentrated in amorphous material within RV replication complexes.
- Unlabeled fine strands were frequently observed alongside the dsRNA-labeled material.
- Detergent treatment released amorphous material from vesicles, suggesting vesicles contain the dsRNA.
- Similar dsRNA-associated structures were found in Semliki Forest virus (SFV) replication complexes.
Conclusions:
- Vesicles lining the replication complexes of both RV and SFV contain viral dsRNA.
- This dsRNA represents viral replicative forms and intermediates.
- The findings elucidate the role of vesicles in housing and releasing viral RNA during replication.