中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2013年
6期
429-431
,共3页
卢明枝%宋娟%孙鹏%宋芹芹%盛琳君%王宝栋%迟苗苗%姚海兰%李朝品
盧明枝%宋娟%孫鵬%宋芹芹%盛琳君%王寶棟%遲苗苗%姚海蘭%李朝品
로명지%송연%손붕%송근근%성림군%왕보동%지묘묘%요해란%리조품
柯萨奇病毒感染%蛋白激酶类%核糖体
柯薩奇病毒感染%蛋白激酶類%覈糖體
가살기병독감염%단백격매류%핵당체
Coxsackievirus infections%Protein kinases%Ribosomes
目的 探讨CVB3的蛋白酶2A对真核细胞帽样蛋白翻译和内部核糖体进入位点(IRES)翻译机制的影响.方法 构建表达载体pcDNA3.1-2A,将此表达载体分别与pEGFP-N1、pGL3(F-luc)以及pIRES-GFP载体共转染细胞后,荧光显微镜观察绿色荧光蛋白(GFP)的表达,检测荧光素酶蛋白的表达.Western Blot检测CVB3病毒和pcDNA3.1-2A对真核生物翻译起始因子4GI(eIF4GI)及多聚A尾结合蛋白(PABP)的影响.结果 pcDNA3.1-2A可以减少帽样依赖的GFP和荧光素酶蛋白的表达,但可以促进IRES依赖的GFP表达.CVB3 2A基因质粒转染和CVB3感染后,均可发现细胞内eIF4G的切割现象;同时CVB3对PABP也有降解作用,而CVB32A基因转染对PABP无明显作用.结论 CVB3的蛋白酶2A抑制真核细胞帽样途径蛋白翻译,促进IRES途径的翻译.
目的 探討CVB3的蛋白酶2A對真覈細胞帽樣蛋白翻譯和內部覈糖體進入位點(IRES)翻譯機製的影響.方法 構建錶達載體pcDNA3.1-2A,將此錶達載體分彆與pEGFP-N1、pGL3(F-luc)以及pIRES-GFP載體共轉染細胞後,熒光顯微鏡觀察綠色熒光蛋白(GFP)的錶達,檢測熒光素酶蛋白的錶達.Western Blot檢測CVB3病毒和pcDNA3.1-2A對真覈生物翻譯起始因子4GI(eIF4GI)及多聚A尾結閤蛋白(PABP)的影響.結果 pcDNA3.1-2A可以減少帽樣依賴的GFP和熒光素酶蛋白的錶達,但可以促進IRES依賴的GFP錶達.CVB3 2A基因質粒轉染和CVB3感染後,均可髮現細胞內eIF4G的切割現象;同時CVB3對PABP也有降解作用,而CVB32A基因轉染對PABP無明顯作用.結論 CVB3的蛋白酶2A抑製真覈細胞帽樣途徑蛋白翻譯,促進IRES途徑的翻譯.
목적 탐토CVB3적단백매2A대진핵세포모양단백번역화내부핵당체진입위점(IRES)번역궤제적영향.방법 구건표체재체pcDNA3.1-2A,장차표체재체분별여pEGFP-N1、pGL3(F-luc)이급pIRES-GFP재체공전염세포후,형광현미경관찰록색형광단백(GFP)적표체,검측형광소매단백적표체.Western Blot검측CVB3병독화pcDNA3.1-2A대진핵생물번역기시인자4GI(eIF4GI)급다취A미결합단백(PABP)적영향.결과 pcDNA3.1-2A가이감소모양의뢰적GFP화형광소매단백적표체,단가이촉진IRES의뢰적GFP표체.CVB3 2A기인질립전염화CVB3감염후,균가발현세포내eIF4G적절할현상;동시CVB3대PABP야유강해작용,이CVB32A기인전염대PABP무명현작용.결론 CVB3적단백매2A억제진핵세포모양도경단백번역,촉진IRES도경적번역.
Objective To study the impact of on eukaryotic cell protein translation by CVB3 2A protease.Methods 293T cells were transfected respectively with plasmids pcDNA3.1-2A,pEGFP-N1,wihch express cap-dependent Green fluoresce protein(GFP) protein or cap-indepent,pGL3 wihch express cap-dependent luciferase protein,pIRES-GFP wihch express cap-independent GFP protein.Green Fluoresce Protein(GFP) luciferase protein were observed and dectedted respectively.The change of eukaryotic translation initiation factor 4G (eIF4G) and the poly (A)-binding protein (PABP) after CVB3 virus infection or CVB3 2A transfectionprotease were detected by Western blotting.Results CVB3 2A inhibited cap-dependent translated both GFP and luciferase protein expression,however encourage the capindependent GFP expression from plamid pIRES-GFP cleavage of eIF4G was detected after both CVB3 infection and CVB3 2A preotease transfection reduction of PABP only was observed after CVB3 infection.Conclusions CVB3 2A protease inhibited of cap-dependent protein translation and encouraged the IRES-dependent protein translation.
