目的:研究汉滩病毒(HTNV)G1蛋白胞质区ITAM样基序与Syk的细胞内相互作用。方法:构建用于研究Syk和G1ITAM样基序之间相互作用的哺乳动物细胞双杂交系统,验证G1ITAM样基序与Syk之间是否存在细胞内相互作用。结果:哺乳动物细胞双杂交分析证实G1ITAM样基序在哺乳动物细胞内可以与Syk相互作用;而突变体分析表明,这种相互作用依赖于该基序中两个高度保守的酪氨酸残基的存在。结论:HTNVG1蛋白胞质区高度保守ITAM样基序在体外和哺乳动物细胞内可以与Syk相互作用,为进一步探讨G1蛋白ITAM样基序在HFRS免疫信号传递中的作用及其与血管内皮损伤的关系奠定了基础。
Objective: To investigate the interactions between Syk and HTNV G1 ITAM-like sequence of Hantaan virus in mammalian ceUs.nethods:A mammalian two-hybrid system was constructed to further confirm interactions between Syk and G1 ITAM-hke-motif in vivo. Results:Mammalian two-hybrid analysis identified that there were direct interactions between the G1 ITAM-like sequence and Syk kinases in the cells. This study showed that two conserved tyrosine residues of ITAM-like sequence in G1 were required in the interactions between this motif and Syk in vivo. Conclusion: The functional ITAM-like sequence in HTNV envelope glycoprotein G1 can bind protein tyrosine kinase Syk which is critical in signal transduction in activation events for immune and endothelial cells. The mimicry of host cell 1TAM in HTNV is a welldemonstrated mechanism through which the virus dysregulates host immune and endothelial cell functions.