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Related Experiment Videos

Insertion of cellular NEDD8 coding sequences in a pestivirus.

M Baroth1, M Orlich, H J Thiel

  • 1Institut für Virologie (FB Veterinärmedizin), Giessen, D-35392, Germany.

Virology
|December 19, 2000
PubMed
Summary

Bovine viral diarrhea virus (BVDV) strain CP 821 contains a novel insertion of cellular protein NEDD8, causing cytopathogenicity. This discovery reveals the genetic basis for BVDV virulence and its phenotypic changes.

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Area of Science:

  • Virology
  • Molecular Biology
  • Genomics

Background:

  • Bovine viral diarrhea virus (BVDV) is a significant pathogen in cattle.
  • The cytopathogenic (cp) strain CP 821 exhibits unique genetic alterations.
  • Understanding viral genetic determinants of pathogenicity is crucial for disease control.

Purpose of the Study:

  • To identify the genetic basis of cytopathogenicity in BVDV strain CP 821.
  • To characterize the nonviral insertion within the BVDV genome.
  • To investigate the role of this insertion in viral replication and phenotype.

Main Methods:

  • Genomic analysis of BVDV CP 821, including molecular cloning and sequencing.
  • Identification of the cellular recombination partner.
  • Introduction of CP 821-specific alterations into a noncytopathogenic (noncp) BVDV clone.

Related Experiment Videos

  • Transfection of bovine cells and recovery of viruses.

    • Main Results:

    • A duplication of viral genes and a nonviral insertion of bovine NEDD8 mRNA were detected in BVDV CP 821.
    • The NEDD8 insertion, coding for a protein 58% identical to ubiquitin, resulted in an additional polyprotein cleavage site.
    • Introduction of these alterations into a noncp BVDV clone generated a cp virus, confirming their role in cytopathogenicity.
    • The study also observed rapid evolution from cp to noncp BVDV phenotypes post-transfection.

    Conclusions:

    • The NEDD8 coding insertion and duplicated viral sequences are the genetic determinants of CP 821 cytopathogenicity.
    • This represents the first report of a viral RNA genome with a NEDD8 coding insertion.
    • The study demonstrates the potential for BVDV to change from cp to noncp phenotype during replication in cell culture.