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Related Experiment Videos

Novel replication complex architecture in rubella replicon-transfected cells.

Juan Fontana1, Wen-Pin Tzeng, Gloria Calderita

  • 1Department of Structure of Macromolecules, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.

Cellular Microbiology
|November 8, 2006
PubMed
Summary
This summary is machine-generated.

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Rubella virus (RUB) replication occurs in modified vacuoles (CPVs). Viral replicase proteins and dsRNA form 2D arrays on CPVs, enabling genome replication on the cytoplasmic side.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Rubella virus (RUB) replication complexes (RCs) assemble in cytopathic vacuoles (CPVs), modified endo-lysosomal organelles.
  • CPVs are crucial RUB replication factories, recruiting cellular components like the endoplasmic reticulum, mitochondria, and Golgi.

Purpose of the Study:

  • To investigate the structure of RUB replication complexes (RCs) within cytopathic vacuoles (CPVs).
  • To determine the role of viral replicase proteins and capsid protein in organelle recruitment and RC formation.

Main Methods:

  • Utilized bicistronic RUB replicons in transfected cells.
  • Employed confocal microscopy to localize viral proteins and dsRNA.
  • Conducted electron microscopy (EM) and immuno-EM to visualize CPV structure and protein localization.

Related Experiment Videos

  • Used streptolysin-O permeabilization for dsRNA labeling.

    • Main Results:

    • RUB replicase components (P90, P150) and dsRNA localized to CPVs.
    • Viral replicase proteins were sufficient for CPV biogenesis and organelle recruitment.
    • CPVs contained protein-rich, sheet-like structures identified as 2D arrays of functional viral replicases.
    • Viral genome replication occurred on the cytoplasmic side of CPVs.
    • Mitochondrial protein P32 was found within CPVs, interacting with the capsid protein (C).

    Conclusions:

    • RUB RCs are organized into 2D arrays of replicase proteins on CPV membranes.
    • Viral replication occurs on the cytoplasmic face of these specialized vacuoles.
    • The capsid protein (C) influences CPV structure, and P32 is a novel component of RUB RCs.