NDV induces ribosomal stress in HeLa cells and affects the translation initiation complex

Abstract:

[Objective]To study the regulation of eIF2α-mediated translation initiation complex eIF4F after Newcastle disease virus (NDV) HBUN/LSRC/F3(hereinafter referred to as NDV F3) induces ribosomal stress in cervical cancer HeLa cells.[Methods]Flow cytometry and cell counting Kit-8(CCK-8) were used to detect cell apoptosis,quantitative real-time polymerase chain reaction to examine c-Myc gene expression,flow cytometry to analyze cell cycle,Western blotting to assess the expression of c-Myc,RPS7,Bcl-2,NP,eIF4E,and eIF2α proteins,and Western blotting and immunofluorescence staining to locate NP and eIF4E proteins.[Results]Compared with the negative control group,NDV F3 inhibited the proliferation of HeLa cells and induced apoptosis.We observed G₀/G₁ arrest,inhibition of c-Myc expression in a time-dependent manner,decrease in protein expression of c-Myc and Bcl-2 in 0-48 h,generation of NP protein at 24 h followed by the increasing trend,and the increase in content of RPS7,eIF4E and eIF2α proteins followed by a decrease during 0-48 h.The results of Western blotting and laser confocal microscopy showed that NP protein was mainly in cytoplasm and that NP co-localized with eIF4E.[Conclusion] NDV F3 induces apoptosis of HeLa cells and triggers ribosomal stress.NP interacts with eIF4E to inhibit the formation of eIF2α-mediated translation initiation complex eIF4F,which blocks the connection with host mRNA and promotes the expression of NDV F3 mRNA,ultimately resulting in the inhibition of host protein translation.