Ainelo Hanna, Lahesaare Andrio, Teppo Annika, Kivisaar Maia, Teras Riho
Chair of Genetics, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
PLoS One. 2017 Sep 25;12(9):e0185482. doi: 10.1371/journal.pone.0185482. eCollection 2017.
LapA is the biggest protein in Pseudomonas putida and a key factor for biofilm formation. Its importance and posttranslational regulation is rather thoroughly studied but less is known about the transcriptional regulation. Here we give evidence that transcription of lapA in LB-grown bacteria is initiated from six promoters, three of which display moderate RpoS-dependence. The global transcription regulator Fis binds to the lapA promoter area at six positions in vitro, and Fis activates the transcription of lapA while overexpressed in cells. Two of the six Fis binding sites, Fis-A7 and Fis-A5, are necessary for the positive effect of Fis on the transcription of lapA in vivo. Our results indicate that Fis binding to the Fis-A7 site increases the level of transcription from the most distal promoter of lapA, whereas Fis binding to the Fis-A5 site could be important for modifying the promoter area topology.
LapA是恶臭假单胞菌中最大的蛋白质,也是生物膜形成的关键因素。其重要性和翻译后调控已得到较为深入的研究,但转录调控方面的了解较少。在此,我们证明在LB培养基中生长的细菌中,lapA的转录起始于六个启动子,其中三个表现出对RpoS的中度依赖性。全局转录调节因子Fis在体外可在六个位置与lapA启动子区域结合,并且在细胞中过表达时Fis可激活lapA的转录。六个Fis结合位点中的两个,即Fis-A7和Fis-A5,对于Fis在体内对lapA转录的正向作用是必需的。我们的结果表明,Fis与Fis-A7位点的结合增加了lapA最远端启动子的转录水平,而Fis与Fis-A5位点的结合对于改变启动子区域拓扑结构可能很重要。