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Genome-Scale CRISPR Screening Reveals Host Factors Required for Ribosome Formation and Viral Replication.

Maikke B Ohlson1, Jennifer L Eitson1, Alexandra I Wells1

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Viruses rely on host cell ribosomes for protein synthesis. This study identified crucial ribosome biogenesis proteins, including SPATA5, as essential host factors for viral replication across diverse virus families.

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Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Viruses hijack host cell machinery, particularly ribosomes, for viral protein synthesis.
  • The specific host factors involved in ribosome biogenesis for viral replication remain largely uncharacterized.

Purpose of the Study:

  • To identify host factors essential for viral protein synthesis and replication using a genome-wide CRISPR screen.
  • To investigate the role of specific ribosome biogenesis proteins, such as SBDS and SPATA5, in the replication of various viruses.

Main Methods:

  • A loss-of-function CRISPR screen was employed to identify host genes required for flavivirus reporter synthesis.
  • Viral replication assays were performed across multiple virus families (Flaviviridae, Coronaviridae, Alphaviruses, Paramyxoviruses, Enterovirus, Poxvirus).
  • Mechanistic studies focused on the function of SPATA5 in rRNA processing and ribosome assembly.

Main Results:

  • Multiple 60S ribosome biogenesis proteins were identified as essential host factors for viral protein synthesis.
  • SBDS and SPATA5 were broadly required for the replication of a wide range of RNA and DNA viruses.
  • Loss of SPATA5 impaired rRNA processing and ribosome assembly, indicating its critical role in ribosome biogenesis.

Conclusions:

  • Specific ribosome biogenesis proteins function as viral host dependency factors, crucial for synthesizing viral proteins and supporting optimal viral replication.
  • SPATA5 is implicated as a key host factor in ribosome formation, potentially acting as a functional ortholog of yeast Drg1.
  • These findings highlight the importance of ribosome biogenesis pathways as targets for antiviral strategies.