Fig. 2: Inhibition of host RNA helicase eIF4A abolishes RSV viral protein synthesis and virus progeny.
From: Human telomerase reverse transcriptase supports respiratory syncytial virus replication
Lenti-viral guide RNAs were used to CRISPR expression of EIF4A. gEIF4A-A549 cells were infected with RSV NH409A (A) or HCoV OC43 (R). A549 cells were infected with clinical isolate NH409A, m.o.i = 0.2 (F), HCoV OC43, m.o.i = 0.5 (T), or influenza H1N1/A/NWS/33, m.o.i = 0.3 (AA) and subsequently treated with varying concentrations of episilvestrol, an EIF4A inhibitor. Viral proteins, RNAs, or progeny was measures at 24 h post infection. A, F Western blot of cellular and RSV viral proteins. Veh vehicle. B–E Quantification of cellular and viral proteins displayed in (A) (n = 3). B EIF4A1/β-actin; C EIF4G1/β-actin; D RSV nucleoprotein N/β-actin; E RSV fusion protein F1/β-actin. Quantification of cellular and viral proteins displayed in (F) (n = 3). G EIF4A1/β-actin; H RSV nucleoprotein N/β-actin; I RSV fusion protein F1/β-actin. J Quantification of virus progeny in the absence and presence of episilvestrol (n = 3). K–Q Quantification of RSV viral RNA expression at presence of indicated concentration of episilvestrol (n = 3). K non-structural protein 2 NS2; L nucleoprotein N; M phosphoprotein P; N matrix protein M; O glycoprotein G; P fusion protein F; Q large polymerase subunit L. R, T Western blot of HCoV viral protein. Veh vehicle. S Quantification of HCoV viral protein spike S displayed in (R) (n = 3). U Quantification of HCoV viral protein S displayed in (T) (n = 3). AA Western blot of influenza viral proteins. Veh vehicle. BB–CC Quantification of viral proteins displayed in panel AA (n = 3). BB influenza nucleoprotein NP/β-actin; CC influenza neuraminidase NA/β-actin. DD–JJ Quantification of influenza viral RNA expression at presence of indicated concentration of episilvestrol (n = 2). DD polymerase basic 1 PB1; EE polymerase acidic PA; FF haemagglutinin HA; GG nucleoprotein NP; HH neuraminidase NA; II matrix protein 1 M1; JJ non-structural protein 1 NS1.Error bars, SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n.s. non-significant.