Engelhardt Institute of Molecular Biology of Russian Academy of Sciences, Moscow, Russia.
Engelhardt Institute of Molecular Biology of Russian Academy of Sciences, Moscow, Russia; Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, Russia.
Redox Biol. 2020 Sep;36:101654. doi: 10.1016/j.redox.2020.101654. Epub 2020 Jul 25.
In recent years, the gasotransmitter hydrogen sulphide (HS), produced by the transsulphuration pathway, has been recognized as a biological mediator playing an important role under normal conditions and in various pathologies in both eukaryotes and prokaryotes. The transsulphuration pathway (TSP) includes the conversion of homocysteine to cysteine following the breakdown of methionine. In Drosophila melanogaster and other eukaryotes, HS is produced by cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulphurtransferase (MST). In the experiments performed in this study, we were able to explore the CRISPR/Cas9 technique to obtain single and double deletions in homozygotes of these three major genes responsible for HS production in Drosophila melanogaster. In most cases, the deletion of one studied gene does not result in the compensatory induction of two other genes responsible for HS production. Transcriptomic studies demonstrated that the deletions of the above CBS and CSE genes alter genome expression to different degrees, with a more pronounced effect being exerted by deletion of the CBS gene. Furthermore, the double deletion of both CBS and CSE resulted in a cumulative effect on transcription in the resulting strains. Overall, we found that the obtained deletions affect numerous genes involved in various biological pathways. Specifically, genes involved in the oxidative reduction process, stress-response genes, housekeeping genes, and genes participating in olfactory and reproduction are among the most strongly affected. Furthermore, characteristic differences in the response to the deletions of the studied genes are apparently organ-specific and have clear-cut sex-specific characteristics. Single and double deletions of the three genes responsible for the production of HS helped to elucidate new aspects of the biological significance of this vital physiological mediator.
近年来，通过转硫途径产生的气体递质硫化氢（HS）已被认为是一种生物介质，在真核生物和原核生物的正常条件下以及各种病理状态下都发挥着重要作用。转硫途径（TSP）包括蛋氨酸分解后同型半胱氨酸转化为半胱氨酸。在黑腹果蝇和其他真核生物中，HS 由胱硫醚β-合酶（CBS）、胱硫醚γ-裂合酶（CSE）和 3-巯基丙酮酸硫转移酶（MST）产生。在本研究中进行的实验中，我们能够探索 CRISPR/Cas9 技术，以获得负责 HS 产生的这三个主要基因在黑腹果蝇中的纯合子的单和双缺失。在大多数情况下，一个研究基因的缺失不会导致两个负责 HS 产生的其他基因的补偿诱导。转录组研究表明，CBS 和 CSE 基因的缺失以不同程度改变基因组表达，CBS 基因的缺失产生更明显的影响。此外，CBS 和 CSE 两个基因的双缺失导致在产生的菌株中转录产生累积效应。总的来说，我们发现获得的缺失会影响涉及各种生物学途径的许多基因。具体而言，参与氧化还原过程、应激反应基因、管家基因以及参与嗅觉和生殖的基因受到的影响最大。此外，研究基因缺失的特征性差异显然是器官特异性的，并且具有明显的性别特异性特征。负责 HS 产生的三个基因的单和双缺失有助于阐明这种重要生理介质的生物学意义的新方面。
