Li Xiao-Dong, Mäkelä Tomi P, Guo Deyin, Soliymani Rabah, Koistinen Vesa, Vapalahti Olli, Vaheri Antti, Lankinen Hilkka
Department of Virology1 and Department of Pathology2, Haartman Institute and HUCH Laboratory Diagnostics, and Institute of Biotechnology3, PO Box 21, Haartmaninkatu 3, FIN-00014 University of Helsinki, Finland.
J Gen Virol. 2002 Apr;83(Pt 4):759-766. doi: 10.1099/0022-1317-83-4-759.
Hantaviruses cause two severe diseases, haemorrhagic fever with renal syndrome in Eurasia and hantavirus pulmonary syndrome in the Americas. To understand more about the molecular mechanisms that lead to these diseases, the associations of Puumala virus nucleocapsid protein (PUUV-N) with cellular proteins were studied by yeast two-hybrid screening. Daxx, known as an apoptosis enhancer, was identified from a HeLa cDNA library and its interaction with PUUV-N was confirmed by GST pull-down assay, co-immunoprecipitation and co-localization studies. Furthermore, domains of interaction were mapped to the carboxyl-terminal region of 142 amino acids in Daxx and the carboxyl-terminal 57 residues in PUUV-N, respectively. In pepscan assays, the binding sites of Daxx to PUUV-N were mapped further to two lysine-rich regions, of which one overlaps the sequence of the predicted nuclear localization signal of Daxx. These data suggest a direct link between host cell machinery and a hantavirus structural component.
汉坦病毒可引发两种严重疾病,在欧亚大陆引发肾综合征出血热,在美洲引发汉坦病毒肺综合征。为了更深入了解导致这些疾病的分子机制,通过酵母双杂交筛选研究了普马拉病毒核衣壳蛋白(PUUV-N)与细胞蛋白的关联。从人宫颈癌HeLa细胞cDNA文库中鉴定出一种名为死亡结构域相关蛋白(Daxx)的凋亡增强因子,并通过谷胱甘肽S-转移酶(GST)下拉试验、免疫共沉淀和共定位研究证实了它与PUUV-N的相互作用。此外,相互作用结构域分别定位于Daxx中142个氨基酸的羧基末端区域和PUUV-N中羧基末端的57个残基。在肽扫描分析中,Daxx与PUUV-N的结合位点进一步定位于两个富含赖氨酸的区域,其中一个区域与Daxx预测的核定位信号序列重叠。这些数据表明宿主细胞机制与汉坦病毒结构成分之间存在直接联系。