Rgg regulates growth phase-dependent expression of proteins associated with secondary metabolism and stress in Streptococcus pyogenes
Rgg regulates growth phase-dependent expression of proteins associated with secondary metabolism and stress in Streptococcus pyogenes
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DOI：
10.1128/jb.186.21.7091-7099.2004
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发表时间：
2004-11-01
影响因子：
3.2
通讯作者：
Chaussee, MS
中科院分区：
文献类型：
--
作者：
Chaussee, MA;Callegari, EA;Chaussee, MS
转录调节蛋白Rgg协调酿脓链球菌中氨基酸催化剂和毒力因子的表达。我们用蛋白质组学方法比较了从S.化脓性链球菌野生型菌株NZ131（血清型M49）与在生长的指数期和稳定期期间从rgg突变菌株分离的蛋白质的差异。蛋白质通过双向凝胶电泳分离，并通过串联质谱鉴定了125个感兴趣的蛋白质点。从同基因菌株分离的蛋白质的比较分析表明，生长阶段相关的监管参与精氨酸（ArcABC），组氨酸（HutI），丝氨酸（SdhA）的代谢酶被废除的RGG突变株，合成的蛋白质在指数生长期。与此相反，酶只检测到野生型蛋白质中分离的生物体在生长的稳定期。蛋白质组成的差异与先前描述的代谢变化相关。此外，与热和氧化应激反应相关的蛋白质，包括ClpE和ClpL，存在于从rgg突变菌株分离的样品中，但不存在于从野生型菌株分离的样品中。rgg突变株比野生型菌株对高温和嘌呤霉素的耐受性更强，但对百草枯的耐受性较差。我们的结论是，RGG是一个全球性的调节因子，有助于生长阶段依赖的蛋白质合成与次生代谢和氧化和热应激反应。
The transcriptional regulatory protein Rgg coordinates amino acid catabolism and virulence factor expression in Streptococcus pyogenes. We used a proteomic approach to compare cytoplasmic proteins isolated from S. pyogenes wild-type strain NZ131 (serotype M49) to proteins isolated from an rgg mutant strain during the exponential and stationary phases of growth. Proteins were separated by two-dimensional gel ellectrophoresis, and 125 protein spots of interest were identified by tandem mass spectrometry. Comparative analysis of proteins isolated from the isogenic strains revealed that growth phase-associated regulation of enzymes involved in the metabolism of arginine (ArcABC), histidine (HutI), and serine (SdhA) was abrogated in the rgg mutant strain, which synthesized the proteins in the exponential phase of growth. In contrast, the enzymes were detected only among wild-type proteins isolated from organisms in the stationary phase of growth. The differences in protein composition were correlated with previously described metabolic changes. In addition, proteins associated with thermal and oxidative stress responses, including ClpE and ClpL, were present in samples isolated from the rgg mutant strain but not in samples isolated from the wild-type strain. The rgg mutant strain was more tolerant to elevated temperature and puromycin than the wild-type strain; however, the mutant was less tolerant to paraquat. We concluded that Rgg is a global regulatory factor that contributes to growth phase-dependent synthesis of proteins associated with secondary metabolism and oxidative and thermal stress responses.