利用全基因组DNA微阵列分析恶臭假单胞菌KT2440转录机制的生长阶段依赖性表达。
Growth phase-dependent expression of the Pseudomonas putida KT2440 transcriptional machinery analysed with a genome-wide DNA microarray.
作者信息
Yuste Luis, Hervás Ana B, Canosa Inés, Tobes Raquel, Jiménez José Ignacio, Nogales Juan, Pérez-Pérez Manuel M, Santero Eduardo, Díaz Eduardo, Ramos Juan-Luis, de Lorenzo Víctor, Rojo Fernando
机构信息
Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus de la Universidad Autónoma de Madrid, Cantoblanco, 28049 - Madrid, Spain.
出版信息
Environ Microbiol. 2006 Jan;8(1):165-77. doi: 10.1111/j.1462-2920.2005.00890.x.
Bacterial transcriptional networks are built on a hierarchy of regulators, on top of which lie the components of the RNA polymerase (in particular the sigma factors) and the global control elements, which play a pivotal role. We have designed a genome-wide oligonucleotide-based DNA microarray for Pseudomonas putida KT2440. In combination with real-time reverse transcription polymerase chain reaction (RT-PCR), we have used it to analyse the expression pattern of the genes encoding the RNA polymerase subunits (the core enzyme and the 24 sigma factors), and various proteins involved in global regulation (Crc, Lrp, Fur, Anr, Fis, CsrA, IHF, HupA, HupB, HupN, BipA and several MvaT-like proteins), during the shift from exponential growth in rich medium into starvation and stress brought about by the entry into stationary phase. Expression of the genes encoding the RNA polymerase core and the vegetative sigma factor decreased in stationary phase, while that of sigma(S) increased. Data obtained for sigma(N), sigma(H), FliA and for the 19 extracytoplasmic function (ECF)-like sigma factors suggested that their mRNA levels change little upon entry into stationary phase. Expression of Crc, BipA, Fis, HupB, HupN and the MvaT-like protein PP3693 decreased in stationary phase, while that of HupA and the MvaT-like protein PP3765 increased significantly. Expression of IHF was indicative of post-transcriptional control. These results provide the first global study of the expression of the transcriptional machinery through the exponential stationary-phase shift in P. putida.
细菌转录网络建立在调控因子的层级之上,RNA聚合酶的组成部分(特别是σ因子)和全局控制元件位于该层级的顶端,它们发挥着关键作用。我们为恶臭假单胞菌KT2440设计了一种基于全基因组寡核苷酸的DNA微阵列。结合实时逆转录聚合酶链反应(RT-PCR),我们用它来分析编码RNA聚合酶亚基(核心酶和24种σ因子)以及参与全局调控的各种蛋白质(Crc、Lrp、Fur、Anr、Fis、CsrA、IHF、HupA、HupB、HupN、BipA和几种MvaT样蛋白)在从富含营养的培养基中的指数生长转变为进入稳定期所带来的饥饿和应激状态过程中的表达模式。编码RNA聚合酶核心和营养型σ因子的基因在稳定期表达下降,而σ(S)的表达增加。关于σ(N)、σ(H)、FliA以及19种胞外功能(ECF)样σ因子的数据表明,进入稳定期后它们的mRNA水平变化不大。Crc、BipA、Fis、HupB、HupN和MvaT样蛋白PP3693的表达在稳定期下降,而HupA和MvaT样蛋白PP3765的表达显著增加。IHF的表达表明存在转录后调控。这些结果首次对恶臭假单胞菌在指数期到稳定期转变过程中转录机制的表达进行了全局研究。