Zakluta A S, Shilova V Y, Zatsepina O G
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.
Mol Biol (Mosk). 2023 Jan-Feb;57(1):139-148. doi: 10.31857/S0026898423010160.
The enzymes involved in the transsulfuration pathway and hydrogen sulfide production-cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (3-MST) - play an important cytoprotective role in the functioning of the organism. Using CRISPER/Cas9 technology, we obtained Drosophila strains with deleted cbs, cse, and mst genes as well as with double deletion of cbs and cse genes. We analyzed the effect of these mutations on the pattern of protein synthesis in the salivary glands of third instar larvae and in the ovaries of mature flies. In the salivary glands of strains with cbs and cse deletions, a decrease was found in the accumulation of the FBP2 storage protein containing 20% methionine amino acid residues. In the ovaries, changes were detected in the level of expression and isofocusing points of proteins involved in cell protection against oxidative stress, hypoxia, and protein degradation. It was shown that in the strains with deletions of transsulfuration enzymes the proteins have a similar degree of oxidation to that of the control strain. A decrease in the total number of proteasomes and their activity was found in the strains with deletions of the cbs and cse genes.
参与转硫途径和硫化氢生成的酶——胱硫醚-β-合酶(CBS)、胱硫醚-γ-裂解酶(CSE)和3-巯基丙酮酸硫转移酶(3-MST)——在生物体的功能发挥中起着重要的细胞保护作用。利用CRISPER/Cas9技术,我们获得了cbs、cse和mst基因缺失以及cbs和cse基因双缺失的果蝇品系。我们分析了这些突变对三龄幼虫唾液腺和成熟果蝇卵巢中蛋白质合成模式的影响。在cbs和cse缺失的品系的唾液腺中,发现含20%甲硫氨酸氨基酸残基的FBP2储存蛋白的积累减少。在卵巢中,检测到参与细胞抗氧化应激、缺氧和蛋白质降解保护的蛋白质的表达水平和等聚焦点发生了变化。结果表明,在转硫酶缺失的品系中,蛋白质的氧化程度与对照品系相似。在cbs和cse基因缺失的品系中,发现蛋白酶体的总数及其活性降低。