Center of Life Sciences, Skolkovo Institute of Science and Technology, Skolkovo, Russia.
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia.
Nucleic Acids Res. 2018 Nov 16;46(20):10810-10826. doi: 10.1093/nar/gky880.
C-proteins control restriction-modification (R-M) systems' genes transcription to ensure sufficient levels of restriction endonuclease to allow protection from foreign DNA while avoiding its modification by excess methyltransferase. Here, we characterize transcription regulation in C-protein dependent R-M system Kpn2I. The Kpn2I restriction endonuclease gene is transcribed from a constitutive, weak promoter, which, atypically, is C-protein independent. Kpn2I C-protein (C.Kpn2I) binds upstream of the strong methyltransferase gene promoter and inhibits it, likely by preventing the interaction of the RNA polymerase sigma subunit with the -35 consensus element. Diminished transcription from the methyltransferase promoter increases transcription from overlapping divergent C-protein gene promoters. All known C-proteins affect transcription initiation from R-M genes promoters. Uniquely, the C.Kpn2I binding site is located within the coding region of its gene. C.Kpn2I acts as a roadblock stalling elongating RNA polymerase and decreasing production of full-length C.Kpn2I mRNA. Mathematical modeling shows that this unusual mode of regulation leads to the same dynamics of accumulation of R-M gene transcripts as observed in systems where C-proteins act at transcription initiation stage only. Bioinformatics analyses suggest that transcription regulation through binding of C.Kpn2I-like proteins within the coding regions of their genes may be widespread.
C 蛋白控制限制修饰(R-M)系统的基因转录,以确保有足够水平的限制内切酶,既能保护自身免受外源 DNA 的侵害,又能避免其被过量甲基转移酶修饰。在这里,我们对 C 蛋白依赖性 R-M 系统 Kpn2I 的转录调控进行了研究。Kpn2I 限制内切酶基因由组成型弱启动子转录,这与 C 蛋白无关,这是非典型的。Kpn2I C 蛋白(C.Kpn2I)结合在强甲基转移酶基因启动子的上游并抑制其转录,可能是通过阻止 RNA 聚合酶 σ 亚基与-35 保守元件的相互作用来实现的。甲基转移酶启动子转录减少会增加重叠的发散 C 蛋白基因启动子的转录。所有已知的 C 蛋白都会影响 R-M 基因启动子的转录起始。独特的是,C.Kpn2I 的结合位点位于其基因的编码区内。C.Kpn2I 作为一种阻碍物,会使延伸的 RNA 聚合酶停滞不前,从而减少全长 C.Kpn2I mRNA 的产生。数学模型表明,这种不同寻常的调节模式导致 R-M 基因转录本积累的动态与仅在转录起始阶段发挥作用的 C 蛋白系统相同。生物信息学分析表明,通过 C.Kpn2I 样蛋白在其基因的编码区内结合进行转录调控可能很普遍。