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Genome-wide transcriptomic and proteomic analysis of the primary response to phosphate limitation in Streptomyces coelicolor M145 and in a DeltaphoP mutant

机译:全基因组的转录组和蛋白质组学分析,对链霉菌M145和DeltaphoP突变体中磷酸盐限制的主要反应

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摘要

Phosphate limitation in Streptomyces and in other bacteria triggers expression changes of a large number of genes. This response is mediated by the two-component PhoR-PhoP system. A Streptomyces coelicolor DeltaphoP mutant (lacking phoP) has been obtained by gene replacement. A genome-wide analysis of the primary response to phosphate limitation using transcriptomic and proteomic studies has been made in the parental S. coelicolor M145 and in the DeltaphoP mutant strains. Statistical analysis of the contrasts between the four sets of data generated (two strains under two phosphate conditions) allowed the classification of all genes into 12 types of profiles. The primary response to phosphate limitation involves upregulation of genes encoding scavenging enzymes needed to obtain phosphate from different phosphorylated organic compounds and overexpression of the high-affinity phosphate transport system pstSCAB. Clear interactions have been found between phosphate metabolism and expression of nitrogen-regulated genes and between phosphate and nitrate respiration genes. PhoP-dependent repressions of antibiotic biosynthesis and of the morphological differentiation genes correlated with the observed DeltaphoP mutant phenotype. Bioinformatic analysis of the presence of PHO boxes (PhoP-binding sequences) in the upstream regions of PhoP-controlled genes were validated by binding of PhoP, as shown by electrophoretic mobility shift assays.
机译:链霉菌和其他细菌中的磷酸盐限制会触发大量基因的表达变化。该响应由两组分PhoR-PhoP系统介导。通过基因置换已经获得了一种链霉菌coelicolor DeltaphoP突变体(缺少phoP)。已经在亲代天蓝色链霉菌M145和DeltaphoP突变株中使用转录组和蛋白质组学研究对磷酸盐限制的主要反应进行了全基因组分析。对所生成的四组数据之间的对比进行统计分析(在两个磷酸盐条件下的两个菌株)可以将所有基因分类为12种类型的谱。对磷酸盐限制的主要反应包括上调编码清除酶的基因,这些清除酶需要从不同的磷酸化有机化合物中获得磷酸盐,并过表达高亲和力的磷酸盐转运系统pstSCAB。在磷酸盐代谢和氮调控基因的表达之间以及磷酸盐和硝酸盐呼吸基因之间发现了明显的相互作用。抗生素生物合成和形态分化基因的PhoP依赖抑制与观察到的DeltaphoP突变表型相关。如电泳迁移率变动分析所示,通过PhoP的结合可以验证PhoP控制基因上游区域中PHO盒(PhoP结合序列)的存在的生物信息学分析。

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