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Host cell proteins interacting with the 3' end of TGEV coronavirus genome influence virus replication

Carmen Galán1, Isabel Sola, Aitor Nogales

  • 1Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, C/Darwin 3, Cantoblanco, 28049 Madrid, Spain.

Virology
|July 8, 2009
PubMed

Insights

Researchers identified cellular proteins crucial for coronavirus RNA synthesis. Silencing poly(A)-binding protein (PABP), hnRNP Q, and glutamyl-prolyl-tRNA synthetase (EPRS) reduced viral RNA, while glyceraldehyde 3-phosphate dehydrogenase (GAPDH) increased it.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Coronavirus RNA synthesis relies on a complex interplay between viral enzymes and host cellular factors.
  • Specific recognition of RNA cis-acting signals at the viral genome ends is essential for replication.
  • Identifying these cellular factors is key to understanding and potentially controlling viral replication.

Purpose of the Study:

  • To identify cellular proteins that interact with the transmissible gastroenteritis coronavirus (TGEV) genome ends.
  • To investigate the functional role of these identified proteins in viral RNA synthesis and replication.

Main Methods:

  • RNA affinity protein purification using TGEV 5' and 3' genome ends as bait.
  • Proteomic analysis to identify bound proteins.
  • Gene silencing experiments using a TGEV replicon and infectious virus to assess protein function.
  • Quantification of viral RNA synthesis post-silencing.

Main Results:

  • Ten cellular proteins were identified, with nine preferentially binding to the 3' end (including PABP, hnRNPs, p100, and aminoacyl-tRNA synthetases) and one to the 5' end (PTB).
  • Silencing of PABP, hnRNP Q, and EPRS led to a 2-3 fold reduction in viral RNA synthesis.
  • Silencing of GAPDH, a control, resulted in a 2-3 fold increase in viral RNA synthesis.

Conclusions:

  • PABP, hnRNP Q, and EPRS positively regulate TGEV replication, likely through interaction with the viral 3' end.
  • GAPDH negatively impacts TGEV infection, suggesting a complex host-pathogen interaction.
  • These findings highlight novel host factors involved in coronavirus replication and potential therapeutic targets.

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