科微学术

微生物学报

NDV诱导HeLa细胞发生核糖体应激及对翻译起始
复合体的影响
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

河北省自然科学基金(H2018405044);河北省高等学校科学技术研究项目(QN2020247);张家口市应用基础研究和人才培养计划(1911038A)


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

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    [目的]研究新城疫病毒(Newcastle disease virus,NDV) HBUN/LSRC/F3株(以下简称NDV F3)诱导宫颈癌细胞(HeLa)发生核糖体应激后对eIF2α介导的翻译起始复合体eIF4F的调控作用。[方法]流式细胞术及CCK-8检测细胞凋亡;实时荧光定量PCR (quantitative real-time polymerase chain reaction,qRT-PCR)检测c-Myc基因表达;流式细胞术分析细胞周期;Western blotting技术检测c-Myc、RPS7、Bcl-2、NP、eIF4E及eIF2α蛋白的表达;Western blotting和免疫荧光染色技术检测NP、eIF4E蛋白定位。[结果]与阴性对照组相比,NDV F3抑制HeLa细胞增殖并诱导细胞凋亡。细胞周期中G0/G1期出现停滞,c-Myc表达呈时间依赖性抑制,c-Myc与Bcl-2蛋白表达量在0-48 h内逐渐下降,NP蛋白在24 h时生成并逐渐增加,RPS7、eIF4E和eIF2α蛋白含量在0-48 h内呈先增加后降低趋势。Western blotting定位分析及激光共聚焦显微镜结果显示NP蛋白主要存在细胞质中,NP与eIF4E存在共定位现象。[结论]NDV F3诱导HeLa细胞凋亡并引发核糖体应激反应,NP与eIF4E相互作用而抑制eIF2α介导的翻译起始复合体eIF4F形成,阻断其与宿主mRNA之间的联系,同时促进NDV F3 mRNA的翻译,最终造成宿主蛋白翻译抑制。

    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.

    参考文献
    相似文献
    引证文献
引用本文

陈琳琳,兰金苹,武彩霞,王静,贾朝霞,曹旭,刘开扬. NDV诱导HeLa细胞发生核糖体应激及对翻译起始
复合体的影响. 微生物学报, 2022, 62(9): 3576-3586

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2022-01-24
  • 最后修改日期:2022-04-11
  • 录用日期:
  • 在线发布日期: 2022-09-05
  • 出版日期: