鼻病毒非结构蛋白2B诱导细胞发生内质网应激
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摘要
为探讨鼻病毒非结构蛋白2B诱导内质网应激和细胞凋亡的机制,本研究构建了鼻病毒非结构蛋白2B的真核表达载体p2B-GFP,通过转染BHK-21细胞检测相关标志蛋白的变化情况。结果显示,非结构蛋白2B定位表达于BHK-21细胞内质网,诱导内质网应激标志蛋白Grp78、CHOP的表达增加,并使活化转录因子6(ATF6)的转录活性增加,还诱导BHK-21细胞发生核浓缩而凋亡,使凋亡标志蛋白PARP发生降解而减少。结果提示,鼻病毒非结构蛋白2B可诱导细胞发生内质网应激,并经该途径诱导细胞凋亡。
To investigate whether the non-structural protein 2Bof rhinovirus 16 induces endoplasmic reticulum stress(ERS)and apoptosis,BHK-21 cells were transfected with a plasmid p2B-GFP expressing rhinovirus 162 Bprotein.Biomarkers of ERS and apoptosis were detected respectively.The results showed that 2Bprotein was localized to the ER apparatus.2Bprotein increased the expressions of ERS marker proteins Grp78 and CHOP and the transcriptional activity of activating transcription factor 6(ATF6).We also found 2Bprotein induced nuclear condensation associated with apoptosis and degradation of marker protein PARP in BHK-21 cells.In conclusion,the rhinovirus 16 2Bprotein induces both ERS and apoptosis.
To investigate whether the non-structural protein 2Bof rhinovirus 16 induces endoplasmic reticulum stress(ERS)and apoptosis,BHK-21 cells were transfected with a plasmid p2B-GFP expressing rhinovirus 162 Bprotein.Biomarkers of ERS and apoptosis were detected respectively.The results showed that 2Bprotein was localized to the ER apparatus.2Bprotein increased the expressions of ERS marker proteins Grp78 and CHOP and the transcriptional activity of activating transcription factor 6(ATF6).We also found 2Bprotein induced nuclear condensation associated with apoptosis and degradation of marker protein PARP in BHK-21 cells.In conclusion,the rhinovirus 16 2Bprotein induces both ERS and apoptosis.
引文
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[10]Zhang HM,Ye X,Su Y,Yuan J,Liu Z,Stein DA,Yang D.Coxsackievirus B3 infection activates the unfolded protein response and induces apoptosis through downregulation of p58IPK and activation of CHOP and SREBP1[J].J Virol,2010,84(17):8446-8459.
[2]Traves SL,Proud D.Viral-associated exacerbations of asthma and COPD[J].Curr Opin Pharmacol,2007,7(3):252-258.
[3]李军,朱启镕.鼻病毒的研究现状[J].中华儿科杂志,2005,43(1):18-20.
[4]Eizirik DL,Cardozo AK,Cnop M.The role for endoplasmic reticulum stress in diabetes mellitus[J].Endocr Rev,2008,29(1):42-61.
[5]Szegezdi E,Logue SE,Gorman AM,Samali A.Mediators of endoplasmic reticulum stress-induced apoptosis[J].EMBO Rep,2006,7(9):880-885.
[6]Mousnier A,Swieboda D,Pinto A,Guedán A,Rogers AV,Walton R,Johnston SL,Solari R.Human rhinovirus 16causes Golgi apparatus fragmentation without blocking protein secretion[J].J Virol,2014,88(20):11671-11685.
[7]de Jong AS,de Mattia F,van Dommelen MM,Lanke K,Melchers WJ,Willems PH,van Kuppeveld FJ.Functional analysis of picornavirus 2B proteins:effects on calcium homeostasis and intracellular protein trafficking[J].J Virol,2008,82(7):3782-3790.
[8]Wang Y,Shen J,Arenzana N,Tirasophon W,Kaufman RJ,Prywes R.Activation of ATF6 and an ATF6 DNA binding site by the endoplasmic reticulum stress response[J].J Biol Chem,2000,275(35):27013-27020.
[9]Weithauser A,Bobbert P,Antoniak S,Bhm A,Rauch BH,Klingel K,Savvatis K,Kroemer HK,Tschope C,Stroux A,Zeichhardt H,Poller W,Mackman N,Schultheiss HP,Rauch U.Protease-activated receptor-2regulates the innate immune response to viral infection in a coxsackievirus B3-induced myocarditis[J].J Am Coll Cardiol,2013,62(19):1737-1745.
[10]Zhang HM,Ye X,Su Y,Yuan J,Liu Z,Stein DA,Yang D.Coxsackievirus B3 infection activates the unfolded protein response and induces apoptosis through downregulation of p58IPK and activation of CHOP and SREBP1[J].J Virol,2010,84(17):8446-8459.