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Distance effects during polyprotein processing in the complementation between defective FMDV RNAs.

Elena Moreno1, Celia Perales2,3,1

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Foot-and-mouth disease virus (FMDV) defective RNAs can complement each other, enabling infection. This study reveals that viral RNA processing involves distance effects, not just protein supply, impacting polyprotein cleavage.

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Area of Science:

  • Virology
  • Molecular Biology
  • Cellular Biology

Background:

  • Foot-and-mouth disease virus (FMDV) is a significant pathogen affecting livestock.
  • Viral RNA replication and protein processing are crucial for FMDV pathogenesis.
  • Defective viral RNAs (dvRNAs) can arise during viral passage and may play roles in viral evolution.

Purpose of the Study:

  • To investigate the mechanism of complementation between FMDV defective RNAs.
  • To elucidate the role of the L protease and 3C protease in polyprotein processing of FMDV.
  • To explore the influence of deleted sequences on viral RNA processing and infectivity.

Main Methods:

  • Passaging FMDV in BHK-21 cells to generate defective RNAs.
  • RNA extraction and characterization of the defective RNA segments.
  • Complementation assays to assess RNA infectivity.
  • Electroporation of L-defective RNA and analysis of protein precursor accumulation.
  • In trans expression of L protein to study its effect on polyprotein processing.

Main Results:

  • FMDV passage yielded two defective RNAs, lacking L or capsid-coding regions, which were infectious via complementation.
  • L-defective RNA electroporation led to P3 precursor accumulation, indicating a dependence on the L sequence.
  • Trans-complementation with L protein restored P3 processing, linking L protease activity to 3C protease-mediated cleavages.
  • Complementation involves more than just supplying L and capsid proteins; distance effects on polyprotein processing are implicated.

Conclusions:

  • Complementation between FMDV defective RNAs is a complex process involving interactions beyond simple protein provision.
  • The L protease's activity is intrinsically linked to the secondary processing events mediated by the 3C protease.
  • Distance effects on polyprotein processing play a significant role in the infectivity and replication of defective viral RNAs.