SNT1与RET之间相互作用的酵母双杂交研究

Study on interaction between suc1-binding neurotrophic target 1 and RET by yeast two-hybrid method

目的:研究suc1-binding neurotrophic target(SNT1)与RET之间的相互作用,寻找RET受体下游的结合蛋白,以深入认识RET下游信号转导机制。方法:将RET胞内域与LexA蛋白融合作为DNA结合蛋白,分别将SNT1、SNT1PTB及SNT1ΔPTB与B42AD蛋白融合作为激活域蛋白,共转化酵母菌后,通过β-半乳糖苷酶活性检测以及氨基酸营养缺陷生长实验分析它们之间的相互作用。结果:SNT1及SNT1PTB与RET之间在酵母中存在结合,而SNT1ΔPTB不与RET结合。结论:证实RET与SNT1之间具有相互作用,SNT1的PTB结构域在介导此相互作用的过程中非常重要。

Objective： To study the interaction between RET （GDNF receptor） with sucl-bingding neurotrophic target （SNT1） and search for the possible downstream substrates or regulatory proteins of RET, so as to reveal the mechanism of downstream signal transduction of RET. Methods： The RET^IC （intracellular domain of RET） was fused to LexA and the product was used as a DNA-binding domain protein. SNT1, SNT1PTB or SNT1APTB （SNT1 without PTB） was separately fused to B42AD and their ptoducts were used as an activation domain protein. The cotransformants were tested by β-galactosidase activity analysis and leucine medium growth analysis was used to study the interaction between SNT1 and RET. Results： Interaction between RET and SNT1 or SNT1PTB was found in yeast, but that between SNT1APTB and RET was not observed. Conclusion： It is confirmed that RET can interact with SNT1 in yeast, and the interaction may be mediated by PTB domain of SNT1.