Noncytotoxic inhibition of viral infection through eIF4F-independent suppression of translation by 4EGi-1.
The eukaryotic initiation factor eIF4F recruits ribosomes to capped mRNAs while eIF2 mediates start codon recognition to initiate protein synthesis. Increasing interest in targeting translation to suppress tumor growth has led to the development of new classes of inhibitors, including 4EGi-1, which disrupts eIF4F complexes. However, the full effects of this inhibitor and its potential uses in the treatment of other disease states remain unclear. Here, we show that overall rates of protein synthesis in primary human cells were affected only modestly by eIF4F disruption using the mTOR inhibitor Torin1, yet were highly sensitive to 4EGi-1. Translational suppression occurred even at concentrations of 4EGi-1 that were below those required to significantly alter eIF4F levels but were instead found to increase the association of ribosomal complexes containing inactive eIF2α. Although highly stable in culture, the effects of 4EGi-1 on both cellular protein synthesis and ribosome association were readily reversible upon inhibitor removal. In addition, despite potently inhibiting translation, prolonged exposure to 4EGi-1 had only modest effects on cell morphology and protein abundance without affecting viability or stress tolerance to any significant degree, although differential effects on heat shock protein (hsp) expression highlighted distinct 4EGi-1-sensitive modes of hsp induction. In contrast, 4EGi-1 potently suppressed poxvirus replication as well as both reactivation and lytic phases of herpesvirus infection. These findings identify a novel way in which 4EGi-1 affects the host cell's protein synthesis machinery and demonstrate its potential as a noncytotoxic inhibitor of diverse forms of viral infection.
National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland.
Robert
R
McMahon
Izabela
I
Zaborowska
Derek
D
Walsh
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology* / Q000633:toxicity; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D039562:Eukaryotic Initiation Factor-4F / Q000037:antagonists & inhibitors; D018259:Herpesvirus 1, Human / Q000187:drug effects*; D006801:Humans; D006835:Hydrazones; D009574:Nitro Compounds / Q000494:pharmacology* / Q000633:toxicity; D014176:Protein Biosynthesis / Q000187:drug effects*; D013844:Thiazoles / Q000494:pharmacology* / Q000633:toxicity; D014616:Vaccinia virus / Q000187:drug effects; D014775:Virus Activation / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
C517852:4EGI-1 compound; D000998:Antiviral Agents; D015852:Eukaryotic Initiation Factor-2; D039562:Eukaryotic Initiation Factor-4F; D006835:Hydrazones; D009574:Nitro Compounds; D013844:Thiazoles