CAJ | 학술논문

番茄丛矮病毒的P19蛋白不仅是一个重要的病毒致病因子,而且还可作为RNA干扰(RNAi)的抑制子.这种作用是通过限制细胞内的小RNA,比如小干扰RNA(siRNAs)和微RNA(miRNAs)来实现.但是目前对P19蛋白在哺乳动物细胞上的作用还未见报道.构建了一株p19稳定表达的293细胞系,即293-p19.流式细胞仪分析发现在293细胞中过量表达P19蛋白可显著引发细胞周期的G2/M阻滞.细胞增殖实验显示,293-p19细胞的DNA复制及细胞生长均受到显著的抑制.此外,研究还发现p19可使人胚肾293细胞内的细胞周期调控子的表达谱发生改变.其中包括上调cyclin A1,CDK2,CDK4,CDK6,p18,cyclin D2,p19INK4d和E2F1,及下调p15,cyclin A,cyclin B1和cyclin E1的表达.上述研究结果提不,p19有可能靶向多个G2/M调控蛋白从而引发细胞的G2/M阻滞.
번가총왜병독적P19단백불부시일개중요적병독치병인자,이차환가작위RNA간우(RNAi)적억제자.저충작용시통과한제세포내적소RNA,비여소간우RNA(siRNAs)화미RNA(miRNAs)래실현.단시목전대P19단백재포유동물세포상적작용환미견보도.구건료일주p19은정표체적293세포계,즉293-p19.류식세포의분석발현재293세포중과량표체P19단백가현저인발세포주기적G2/M조체.세포증식실험현시,293-p19세포적DNA복제급세포생장균수도현저적억제.차외,연구환발현p19가사인배신293세포내적세포주기조공자적표체보발생개변.기중포괄상조cyclin A1,CDK2,CDK4,CDK6,p18,cyclin D2,p19INK4d화E2F1,급하조p15,cyclin A,cyclin B1화cyclin E1적표체.상술연구결과제불,p19유가능파향다개G2/M조공단백종이인발세포적G2/M조체.
Besides its function as a pathogenicity determinant, the Tombusvirus P19 also serves as a suppressor of RNA interference (RNAi) by sequestering intracellular small RNAs such as the small interfering RNAs (siRNAs) and microRNAs (miRNAs). However, the effect of P19 on mammalian cells has not been evaluated before. A human embryonic kidney 293 cell line that stably expressed p19 (HEK293-p19) was generated. Flow cytometric analysis revealed that over-expression of P19 caused a significant accumulation of G2/M phase cells. Cell proliferation assays demonstrated a reduced DNA replication and cell growth in HEK293-p19 cells. Moreover, p19 altered the expression profiles of a number of cell cycle regulators in HEK293 cells, such as upregulafion of cyclin A1, CDK2, CDK4, CDK6, p18, cyclin D2, p19INK4d and E2F1, and downregulation of p15, cyclin A2, cyclin B1 and cyclin E1. Thus, the data strongly indicate that p19 might influence multiple G2/M regulators to cause G2/M arrest.