Kawalek Adam, Glabski Krzysztof, Bartosik Aneta Agnieszka, Fogtman Anna, Jagura-Burdzy Grazyna
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Department of Microbial Biochemistry, Warsaw, Poland.
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Laboratory of Microarray Analysis, Warsaw, Poland.
PLoS One. 2017 Jul 21;12(7):e0181726. doi: 10.1371/journal.pone.0181726. eCollection 2017.
Similarly to its homologs in other bacteria, Pseudomonas aeruginosa partitioning protein ParB facilitates segregation of newly replicated chromosomes. Lack of ParB is not lethal but results in increased frequency of anucleate cells production, longer division time, cell elongation, altered colony morphology and defective swarming and swimming motility. Unlike in other bacteria, inactivation of parB leads to major changes of the transcriptome, suggesting that, directly or indirectly, ParB plays a role in regulation of gene expression in this organism. ParB overproduction affects growth rate, cell division and motility in a similar way as ParB deficiency. To identify primary ParB targets, here we analysed the impact of a slight increase in ParB level on P. aeruginosa transcriptome. ParB excess, which does not cause changes in growth rate and chromosome segregation, significantly alters the expression of 176 loci. Most notably, the mRNA level of genes adjacent to high affinity ParB binding sites parS1-4 close to oriC is reduced. Conversely, in cells lacking either parB or functional parS sequences the orfs adjacent to parS3 and parS4 are upregulated, indicating that direct ParB- parS3/parS4 interactions repress the transcription in this region. In addition, increased ParB level brings about repression or activation of numerous genes including several transcriptional regulators involved in SOS response, virulence and adaptation. Overall, our data support the role of partitioning protein ParB as a transcriptional regulator in Pseudomonas aeruginosa.
与其他细菌中的同源物类似,铜绿假单胞菌分区蛋白ParB促进新复制染色体的分离。缺乏ParB并不致命,但会导致无核细胞产生频率增加、分裂时间延长、细胞伸长、菌落形态改变以及群体游动和游泳运动缺陷。与其他细菌不同,parB的失活会导致转录组发生重大变化,这表明ParB直接或间接地在该生物体的基因表达调控中发挥作用。ParB的过量表达对生长速率、细胞分裂和运动性的影响与ParB缺乏类似。为了确定ParB的主要靶标,我们在此分析了ParB水平的轻微增加对铜绿假单胞菌转录组的影响。ParB过量表达不会导致生长速率和染色体分离的变化,但会显著改变176个基因座的表达。最值得注意的是,靠近oriC的高亲和力ParB结合位点parS1 - 4附近的基因的mRNA水平降低。相反,在缺乏parB或功能性parS序列的细胞中,与parS3和parS4相邻的开放阅读框上调,表明直接的ParB - parS3/parS4相互作用抑制了该区域的转录。此外,ParB水平的增加会导致许多基因的抑制或激活,包括参与SOS反应、毒力和适应性的几种转录调节因子。总体而言,我们的数据支持分区蛋白ParB作为铜绿假单胞菌转录调节因子的作用。