粗糙脉孢菌转录因子LAH-3参与渗透压调控

LAH-3，a transcriptional factor involved in osmotic regulation in Neurospora crassa

摘要:

摘要:【目的】通过构建转录因子lah-3基因缺失突变体，研究lah-3基因缺失突变体菌株的渗透压表型，进而探究lah-3基因在渗透压调控中的作用。【方法】采用同源基因重组敲除技术构建lah-3基因缺失突变体。用4% NaCl和1 mol/L Sorbitol进行渗透压处理。利用Northern blot检测渗透压应答基因的表达。利用Westhern blot检测LAH-3 蛋白磷酸化修饰水平，OS-2蛋白的表达水平及其磷酸化修饰水平。【结果】在转录因子lah-3基因缺失突变体中，渗透压应答基因gcy-1、stl-1 以及pck-1的表达水平都明显降低，而且在渗透压刺激下，LAH-3蛋白磷酸化修饰水平升高。LAH-3的磷酸化修饰不受OS-2调控。lah-3基因的缺失既不影响OS-2蛋白的表达水平，也不影响其在渗透压刺激后的磷酸化修饰。【结论】粗糙脉孢菌中转录因子LAH-3参与调控渗透压应答基因的转录，但其响应过程不依赖于OS-2 MAPK信号通路。

Abstract:

Abstract:［Objective］In order to identify the function of lah-3 in osmotic regulation，we generated lah-3 deletion strain and analyzed its phenotype by osmostress treatment．［Methods］We used homologous recombination to replace lah-3gene by hph gene and treated these cells with 4% NaCl and 1 M sorbitol to analyze the phenotype．Northern blot was used to detect the expressions of osmoresponsing genes．Western blot was used to examine the phosphorylation level of LAH-3 and OS-2 and the expression of OS-2．［Results］In the deletion strain of transcription factor lah-3 gene，the expressions of osmoresponsing genes gcy-1，stl-1 and pck-1 were significantly reduced． Besides，the phosphorylation level of LAH-3 protein increased under the osmostress treatment． The phosphorylation of LAH-3 was not mediated by OS-2．The deletion of lah-3 did not affect the os-2 expression and the phosphorylation level of OS-2 upon osmostress．［Conclusion］LAH-3 involved in osmoresponsing was independent of OS-2 MAPK pathway．