酿酒酵母Snf1/AMPK蛋白激酶通过调节细胞壁合成相关基因的表达影响细胞壁完整性

Snf1/AMPK affects cell wall integrity through regulating the transcription of cell wall as sembly-related genesin Saccharomyces cerevisiae

摘要:

[目的] 初步探讨酿酒酵母(Saccharomyces cerevisiae)中Snf1/AMPK蛋白激酶影响细胞壁完整性的机制。[方法] 通过同源重组交换的方法,构建酿酒酵母Snf1/AMPK蛋白激酶催化亚基的敲除菌株snf1Δ,并通过基因回补对敲除菌株表型进行验证。在含有刚果红(Congo red)和荧光增白剂(Calcofluorwhite)的平板上检测snf1Δ菌株细胞壁的完整性,通过qRT-PCR的方法检测snf1Δ菌株中已知的细胞壁合成相关基因的表达情况。[结果] SNF1基因敲除影响细胞壁的完整性,并影响酿酒酵母对热激应答的反应。进一步研究发现,SNF1突变菌株中β-1, 3-葡聚糖合成相关基因与β-1, 6-葡聚糖合成相关基因的表达量均明显降低。[结论] 结果显示酿酒酵母Snf1蛋白激酶影响细胞壁的完整性,此影响发生在转录水平上,即通过调节细胞壁合成相关基因的转录来实现,揭示了Snf1蛋白的一个新角色。

Abstract:

[Objective] To study the role of Snf1/AMPK kinase in cell wall integrity in Saccharomyces cerevisiae JY102. [Methods] SNF1 deletion mutant of S. cerevisiae was created by homologous recombination. Congo red and Calcofluor white were applied to evaluate the cell wall integrity for the mutant strain. qRT-PCR was used to analyze the transcription of cell wall-related genes. [Results] The SNF1 disruption mutant was severely sensitive to Congo red and Calcofluor white, manifesting the impairment of cell wall integrity. The mutant exhibited apparently growth defect at high temperature. The results of qRT-PCR revealed down-regulated transcription of several genes involved in β-glucan biosynthesis. [Conclusion] The deletion of Snf1 impairs the cell wall integrity by reducing the transcription of β-glucan-related genes, suggesting a new role of Snf1 in the activation of cell wall synthesis.