Malesevic Jelena, Kojic Milorad, Stanovcic Stefan, Azanjac Natalija, Milisavljevic Mira
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042 Belgrade, Serbia.
J Fungi (Basel). 2022 Sep 13;8(9):957. doi: 10.3390/jof8090957.
Much headway has been made in understanding the numerous strategies that enable microorganisms to counteract various types of environmental stress, but little is known about how microbial populations recover after a massive death caused by exposure to extreme conditions. Using the yeast-like fungus as a model, our recent post-stress regrowth under starvation (RUS) studies have demonstrated that this organism reconstitutes devastated populations with remarkable efficiency. Subsequently, we have identified four RUS-gene products. Two of these, Did4 and Tbp1, play parallel roles in protecting the genome. To identify additional molecular components, we took a molecular-genetic and a transcriptomic approach. By employing a simple and novel screening method, we identified five RUS-deficient mutants (, , , , ), three of which (, , and ) displayed sensitivity to different genotoxic agents, indicating that the corresponding gene products have roles in genome protection. The global transcriptomic changes of cells grown in supernatants derived from peroxide-treated cell suspensions revealed sets of uniquely expressed genes. Importantly, among the genes induced by the substrates was , which encodes a protein kinase required for checkpoint-mediated cell cycle arrest in response to DNA damage. Mutants of deleted of are severely incapacitated in RUS.
在理解使微生物能够应对各种环境压力的众多策略方面已经取得了很大进展,但对于微生物群体在因暴露于极端条件而导致大量死亡后如何恢复却知之甚少。以酵母样真菌为模型,我们最近关于饥饿后应激再生(RUS)的研究表明,这种生物体能够以显著的效率重建遭受破坏的群体。随后,我们鉴定出了四种RUS基因产物。其中两种,Did4和Tbp1,在保护基因组方面发挥着平行作用。为了鉴定其他分子成分,我们采用了分子遗传学和转录组学方法。通过使用一种简单而新颖的筛选方法,我们鉴定出了五个RUS缺陷突变体(,,,,),其中三个(,和)对不同的基因毒性剂表现出敏感性,这表明相应的基因产物在基因组保护中发挥作用。在过氧化物处理的细胞悬液上清中生长的细胞的全局转录组变化揭示了一组独特表达的基因。重要的是,在由底物诱导的基因中,是,它编码一种在响应DNA损伤时介导检查点介导的细胞周期停滞所需的蛋白激酶。缺失的突变体在RUS中严重丧失能力。