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Related Experiment Videos

Membrane targeting sequences in tombusvirus infections

L Rubino1, M Russo

  • 1Dipartimento di Protezione delle Piante, Università degli Studi, Bari, Italy. csvvlr02@area.ba.cnr.it

Virology
|January 8, 1999
PubMed
Summary

Plant viruses Cymbidium ringspot virus (CymRSV) and Carnation Italian ringspot virus (CIRV) induce membrane-bound compartments. Viral protein sequences dictate whether peroxisomes or mitochondria are modified to form these structures.

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Area of Science:

  • Plant Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Infection by Cymbidium ringspot virus (CymRSV) and Carnation Italian ringspot virus (CIRV) in Nicotiana benthamiana cells leads to the formation of distinct membranous structures called multivesicular bodies (MVBs).
  • These MVBs originate from modified peroxisomes (for CymRSV) or mitochondria (for CIRV).

Purpose of the Study:

  • To investigate the role of viral proteins in directing the modification of host organelles for MVB formation.
  • To identify the specific protein domains responsible for targeting peroxisomes or mitochondria.

Main Methods:

  • Construction and analysis of hybrid viral genomes by exchanging sequences between CymRSV and CIRV ORF 1.
  • In vitro synthesis of viral RNA and inoculation of Nicotiana benthamiana plants and protoplasts.

Related Experiment Videos

  • Biochemical analysis of viral proteins and mutational analysis of the CIRV 36-kDa protein.

    • Main Results:

    • The 33 kDa (CymRSV) and 36 kDa (CIRV) proteins encoded by ORF 1 are integral membrane proteins.
    • Both the N-terminal hydrophilic region and transmembrane segments of these viral proteins determine organelle specificity for MVB synthesis.
    • The CIRV 36-kDa protein contains a mitochondrial targeting sequence, while the CymRSV 33-kDa protein lacks typical peroxisomal targeting signals.

    Conclusions:

    • Viral protein structure dictates the host organelle (peroxisome or mitochondria) modified for MVB formation during infection.
    • The N-terminal region and transmembrane domains of ORF 1 proteins are crucial for organelle targeting.
    • CymRSV and CIRV employ distinct strategies involving their ORF 1 proteins to manipulate host cell compartments.