人博卡病毒HBoV1非结构蛋白NP1对细胞转录因子的调控  

Modulation of cellular transcriptional factors by human bocavirus 1 nonstructural protein NP1

作　　者：孙彬[1] 李永淑[1] 董衍明[1] 刘凯于[1] 杨勇波[1] 李毅[1,2] 

机构地区：[1]华中师范大学生命科学学院,武汉430079 [2]武汉生物工程学院生物工程系,武汉430415

出　　处：《微生物学报》2013年第7期737-745,共9页Acta Microbiologica Sinica

基　　金：国家自然科学基金项目(30670081)~~

摘　　要：【目的】研究人博卡病毒非结构蛋白NP1对细胞转录因子活性和炎症细胞因子TNF-α和IL-6表达的调控作用。【方法】通过双荧光素酶报告基因系统分析NP1对转录因子的调控作用,通过ELISA和荧光定量PCR分别从蛋白质和RNA水平检测NP1是否影响TNF-α、IL-6的表达,最后利用哺乳动物双杂交系统分析NP1是否通过寡聚化发挥功能。【结果】在293T细胞中,NP1可分别提高AP-1、STAT3和GAS转录因子报告载体荧光素酶活性3.69倍、2.45倍和3.03倍,但对NF-κB转录因子报告载体荧光素酶活性无明显影响(P>0.05)。转染24 h和48 h后,NP1表达细胞和对照细胞培养基中IL-6和TNF-α蛋白浓度没有显著性差异(P>0.05),但TNF-αmRNA的表达水平上调。哺乳动物双杂交实验表明NP1蛋白自身没有相互作用。【结论】结果首次表明HBoV1非结构蛋白NP1对细胞转录因子和炎症细胞因子具有调节作用,揭示NP1可能与HBoV1致病性有关,为进一步探究HBoV1病毒致病的分子机制提供参考。[ Objective] We studied the regulating effect of human bocavirus 1 （HBoV1） nonstructural protein NP1 on the activity of cellular transcription factors and the expression of inflammatory cytokine TNF-α and IL-6. [ Methods] The modulation of NP1 was measured by the Dual Luciferase Reporter Assay System and the expression of cytokines TNF-α and IL-6 was detected by ELISA and Real-time PCR. The lueiferase based mammalian two-hybrid system was used to analyze whether the function of NP1 protein aroused from oligomerization. [ Results] The transcription factors AP-1, STAT3 and STAT1 but not NF-KB were up-regulated by NP1, which was evidenced by approximately 2-3-fold increase of the luciferase activity compared to the control vector. Moreover, NP1 increased the TNF-α mRNA expression, but not contributed to cytokine IL-6 secretion. We also found that the self-interaction did not exist when NP1 was solely expressed in 293T cells. [ Conclusionl This study demonstrates for the first time that NP1 may play important roles in activation of transcription factors and up-regulation of inflammatory cytokines expression, suggesting that NP1 be involved in HBoV1 pathogenesis.

关 键 词：人博卡病毒 非结构蛋白NP1 转录因子 细胞因子 

分 类 号：R373[医药卫生—病原生物学]