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Epstein-Barr Virus BGLF2 commandeers RISC to interfere with cellular miRNA function.

Ashley M Campbell1, Carlos F De La Cruz-Herrera1, Edyta Marcon2

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Epstein-Barr virus BGLF2 protein disrupts miRNA regulation by interacting with RISC, leading to increased SUMOylation. This novel function impacts cellular processes and viral infection outcomes.

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Area of Science:

  • Virology
  • Molecular Biology
  • Post-transcriptional Gene Regulation

Background:

  • Epstein-Barr virus (EBV) BGLF2 protein is known to induce SUMOylation.
  • MicroRNAs (miRNAs) regulate gene expression post-transcriptionally via the miRNA-Induced Silencing Complex (RISC).

Purpose of the Study:

  • To investigate the interaction between EBV BGLF2 protein and RISC.
  • To elucidate the functional consequences of BGLF2 interaction with RISC on miRNA activity and SUMOylation.

Main Methods:

  • Affinity-purification coupled to mass-spectrometry to identify BGLF2 interactors.
  • Co-immunoprecipitation and immunofluorescence to confirm protein interactions and localization.
  • Reporter assays to assess miRNA activity.
  • Western blotting to measure protein levels.

Main Results:

  • BGLF2 directly interacts with RISC components Ago2 and TNRC6, co-localizing in cytoplasmic bodies.
  • BGLF2 expression disrupts processing bodies and inhibits let-7 miRNA activity, affecting SUMO transcript regulation.
  • BGLF2 increases cellular unconjugated SUMO proteins, driving SUMOylation and inhibiting miRNA loading into Ago2.

Conclusions:

  • EBV BGLF2 protein functions as a novel miRNA regulator by interfering with RISC.
  • BGLF2 dysregulates SUMO transcripts, leading to increased free SUMO proteins and cellular SUMOylation.
  • This mechanism contributes to EBV's effects on cellular processes during infection.