Skryabin Institute of Biochemistry and Physiology of Microorganisms, FRC Pushchino Center for Biological Research of the Russian Academy of Sciences, pr. Nauki 5, Pushchino 142290, Russia.
Institute for Information Transmission Problems, Russian Academy of Sciences, Bolshoy Karetny per. 19 bld .1, Moscow 127051, Russia.
Cells. 2019 May 15;8(5):461. doi: 10.3390/cells8050461.
Inorganic polyphosphate (polyP) is crucial for adaptive reactions and stress response in microorganisms. A convenient model to study the role of polyP in yeast is the strain CRN/PPN1 that overexpresses polyphosphatase Ppn1 with stably decreased polyphosphate level. In this study, we combined the whole-transcriptome sequencing, fluorescence microscopy, and polyP quantification to characterize the CRN/PPN1 response to manganese and oxidative stresses. CRN/PPN1 exhibits enhanced resistance to manganese and peroxide due to its pre-adaptive state observed in normal conditions. The pre-adaptive state is characterized by up-regulated genes involved in response to an external stimulus, plasma membrane organization, and oxidation/reduction. The transcriptome-wide data allowed the identification of particular genes crucial for overcoming the manganese excess. The key gene responsible for manganese resistance is encoding a low-affinity manganese transporter: Strong down-regulation in CRN/PPN1 increases manganese resistance by reduced manganese uptake. On the contrary, , the top up-regulated gene in CRN/PPN1, is also strongly up-regulated in the manganese-adapted parent strain. Phm7 is an unannotated protein, but manganese adaptation is significantly impaired in Δ, thus suggesting its essential function in manganese or phosphate transport.
无机多聚磷酸盐(polyP)对微生物的适应性反应和应激反应至关重要。研究酵母中多聚磷酸盐作用的一个方便模型是过表达多聚磷酸酶 Ppn1 并使其多聚磷酸盐水平稳定降低的 CRN/PPN1 菌株。在这项研究中,我们结合全转录组测序、荧光显微镜和多聚磷酸盐定量来描述 CRN/PPN1 对锰和氧化应激的反应。由于在正常条件下观察到的预适应状态,CRN/PPN1 对锰和过氧化物表现出增强的抗性。预适应状态的特征是参与对外界刺激、质膜组织和氧化/还原反应的上调基因。全转录组数据允许鉴定克服锰过量的关键基因。负责锰抗性的关键基因是 编码低亲和力锰转运蛋白:在 CRN/PPN1 中的强烈下调通过减少锰摄取来增加锰抗性。相反, 在 CRN/PPN1 中上调最多的基因,在锰适应的亲本菌株中也被强烈上调。Phm7 是一个未注释的蛋白质,但在 Δ 中,锰适应显著受损,这表明它在锰或磷酸盐运输中具有重要功能。