Dissecting regulatory networks by means of two-dimensional gel electrophoresis:: Application to the study of the diauxic shift in the yeast Saccharomyces cerevisiae
Dissecting regulatory networks by means of two-dimensional gel electrophoresis:: Application to the study of the diauxic shift in the yeast Saccharomyces cerevisiae
复制标题
DOI：
10.1002/pmic.200300564
复制
发表时间：
2004-02-01
期刊：
影响因子：
3.4
通讯作者：
Boucherie, H
中科院分区：
文献类型：
--
作者：
Haurie, V;Sagliocco, F;Boucherie, H
使用蛋白质组学方法的基础上的二维（2-D）凝胶分析合成的蛋白质，我们研究了参与的Snf 1激酶途径在调控基因表达在酿酒酵母的二倍性转变。为此，我们使用了缺失SNF 4（编码Snf 1 p激活子亚基的基因）的突变株。82个斑点的合成水平被发现受到Snf 4p在二次移位的情况下。在突变株中表现出合成减少的蛋白质中有一半是其基因受转录激活因子Cat 8 p（Snf 1 p的靶点）控制的蛋白质。还观察到突变株中合成水平增加的蛋白质。其中有糖酵解酶，当野生型细胞进入二次转换时，糖酵解酶的合成强烈减少。这一观察结果表明，Snf 1 p发挥负控制糖酵解基因的表达在二次发育过渡。在这项研究中获得的结果与以前获得的类似的蛋白质组学方法与其他调控因子参与的二次移位。这个汇编说明了如何2-D凝胶电泳可以用来阐明参与复杂的生物过程的调控网络。
Using a proteomic approach based on the two-dimensional (2-D) gel analysis of synthesized proteins, we investigated the involvement of the Snf1 kinase pathway in the regulation of gene expression during the diauxic shift in Saccharomyces cerevisiae. For this purpose, we used a mutant strain deleted for SNF4, the gene coding for the activator subunit of Snf1p. The levels of synthesis of 82 spots were found to be affected by the absence of Snf4p at the diauxic shift. Half of the proteins which exhibit a reduced synthesis in the mutant strain are proteins whose genes are controlled by the transcriptional activator Cat8p, a target of Snf1p. Proteins with an increased level of synthesis in the mutant strain were also observed. Among them are glycolytic enzymes whose synthesis is strongly reduced when wild-type cells enter the diauxic shift. This observation suggests that Snf1p exerts a negative control on the expression of glycolytic genes during the diauxic transition. The results obtained in this study were compiled with those previously obtained by similar proteomic approach with other regulatory factors involved in the diauxic shift. This compilation illustrates how 2-D gel electrophoresis can be used to elucidate the network of regulators participating to complex biological process.