Pletnev Philipp, Guseva Ekaterina, Zanina Anna, Evfratov Sergey, Dzama Margarita, Treshin Vsevolod, Pogorel'skaya Alexandra, Osterman Ilya, Golovina Anna, Rubtsova Maria, Serebryakova Marina, Pobeguts Olga V, Govorun Vadim M, Bogdanov Alexey A, Dontsova Olga A, Sergiev Petr V
Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow, Russia.
Front Genet. 2020 Feb 27;11:97. doi: 10.3389/fgene.2020.00097. eCollection 2020.
Ribosomal RNAs in all organisms are methylated. The functional role of the majority of modified nucleotides is unknown. We systematically questioned the influence of rRNA methylation in on a number of characteristics of bacterial cells with the help of a set of rRNA methyltransferase (MT) gene knockout strains from the Keio collection. Analysis of ribosomal subunits sedimentation profiles of the knockout strains revealed a surprisingly small number of rRNA MT that significantly affected ribosome assembly. Accumulation of the assembly intermediates was observed only for the knockout strain whose growth was retarded most significantly among other rRNA MT knockout strains. Accumulation of the 17S rRNA precursor was observed for () knockout cells as well as for cells devoid of functional and genes. Significant differences were found among the WT and the majority of rRNA MT knockout strains in their ability to sustain exogenous protein overexpression. While the majority of the rRNA MT knockout strains supported suboptimal reporter gene expression, the strain devoid of the gene demonstrated a moderate increase in the yield of ectopic gene expression. Comparative 2D protein gel analysis of rRNA MT knockout strains revealed only minor perturbations of the proteome.
所有生物体中的核糖体RNA都会发生甲基化。大多数修饰核苷酸的功能作用尚不清楚。我们借助来自Keio文库的一组核糖体RNA甲基转移酶(MT)基因敲除菌株,系统地研究了核糖体RNA甲基化对细菌细胞多种特性的影响。对敲除菌株核糖体亚基沉降图谱的分析显示,显著影响核糖体组装的核糖体RNA甲基转移酶数量惊人地少。仅在敲除菌株中观察到组装中间体的积累,该菌株在其他核糖体RNA甲基转移酶敲除菌株中生长受抑制最为显著。在()敲除细胞以及缺乏功能性和基因的细胞中观察到17S核糖体RNA前体的积累。在野生型和大多数核糖体RNA甲基转移酶敲除菌株维持外源蛋白过表达的能力方面发现了显著差异。虽然大多数核糖体RNA甲基转移酶敲除菌株支持次优的报告基因表达,但缺乏基因的菌株在异位基因表达产量上有适度增加。对核糖体RNA甲基转移酶敲除菌株的二维蛋白质凝胶比较分析仅显示蛋白质组有轻微扰动。