Sulfur-limitation-regulated proteins in Bacillus subtilis:: a two-dimensional gel electrophoresis study
Sulfur-limitation-regulated proteins in Bacillus subtilis:: a two-dimensional gel electrophoresis study
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DOI：
10.1099/00221287-147-6-1631
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发表时间：
2001-06-01
期刊：
影响因子：
2.8
通讯作者：
Martin-Verstraete, I
中科院分区：
文献类型：
--
作者：
Coppée, JY;Auger, S;Martin-Verstraete, I
关于枯草芽孢杆菌中参与硫同化的基因和酶，或者关于它们的表达或活性的调节，知之甚少。为了鉴定受硫限制调控的基因，作者使用二维（2D）凝胶电泳比较了以硫酸盐或谷胱甘肽作为唯一硫源生长的野生型菌株的蛋白质组。通过基质辅助激光解吸/电离飞行时间（MALDI TOF）质谱法鉴定了在这两种条件下合成被修饰的总共15种蛋白质。在硫酸盐存在下，观察到参与C1单位（SerA、GlyA、FolD）代谢以及嘌呤（PurQ、Xpt）和嘧啶（Upp、PyrAA、PyrF）生物合成的蛋白质的量增加。在谷胱甘肽的存在下，两个摄取系统（DppE，SsuA），加氧酶（SsuD），半胱氨酸合酶（CysK）和两个未知功能的蛋白质（Ytml，YurL）的合成增加。在硫酸盐和谷胱甘肽的存在下，使用狭缝印迹分析和lacZ融合来监测相应基因的表达的变化。ytml基因是响应于硫可用性而共调节的12个基因的基因座的一部分。这种推定的操纵子由LysR样调节子YtII激活。这是在枯草芽孢杆菌中鉴定的第一种参与响应硫可用性的表达控制的调节子。
Little is known about the genes and enzymes involved in sulfur assimilation in Bacillus subtilis, or about the regulation of their expression or activity. To identify genes regulated by sulfur limitation, the authors used two-dimensional (2D) gel electrophoresis to compare the proteome of a wild-type strain grown with either sulfate or glutathione as sole sulfur source. A total of 15 proteins whose synthesis is modified under these two conditions were identified by matrix-assisted laser desorption/ionization time of flight (MALDI TOF) mass spectrometry, In the presence of sulfate, an increased amount of proteins involved in the metabolism of C, units (SerA, GlyA, FolD) and in the biosynthesis of purines (PurQ, Xpt) and pyrimidines (Upp, PyrAA, PyrF) was observed. In the presence of glutathione, the synthesis of two uptake systems (DppE, SsuA), an oxygenase (SsuD), cysteine synthase (CysK) and two proteins of unknown function (Ytml, YurL) was increased. The changes in expression of the corresponding genes, in the presence of sulfate and glutathione, were monitored using slot-blot analyses and lacZ fusions, The ytml gene is part of a locus of 12 genes which are co-regulated in response to sulfur availability. This putative operon is activated by a LysR-like regulator, Ytll, This is the first regulator involved in the control of expression in response to sulfur availability to be identified in a, subtilis.