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

摘要:

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-α mRNA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。

Abstract:

Abstract:［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-αand IL-6．［Methods］ The modulation of NP1 was measured by the Dual Luciferase Reporter Assay System and the expression of cytokines TNF-αand IL-6 was detected by ELISA and Real-time PCR． The luciferase 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-κB 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．［Conclusion］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．