Skirball Institute of Biomolecular Medicine, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
Department of Microbiology, New York University School of Medicine, 430 E. 29th Street, New York, NY 10016, USA.
Nucleic Acids Res. 2021 Nov 8;49(19):11038-11049. doi: 10.1093/nar/gkab884.
A key pathway for mRNA degradation in bacterial cells begins with conversion of the initial 5'-terminal triphosphate to a monophosphate, a modification that renders transcripts more vulnerable to attack by ribonucleases whose affinity for monophosphorylated 5' ends potentiates their catalytic efficacy. In Escherichia coli, the only proteins known to be important for controlling degradation via this pathway are the RNA pyrophosphohydrolase RppH, its heteromeric partner DapF, and the 5'-monophosphate-assisted endonucleases RNase E and RNase G. We have now identified the metabolic enzyme cytidylate kinase as another protein that affects rates of 5'-end-dependent mRNA degradation in E. coli. It does so by utilizing two distinct mechanisms to influence the 5'-terminal phosphorylation state of RNA, each dependent on the catalytic activity of cytidylate kinase and not its mere presence in cells. First, this enzyme acts in conjunction with DapF to stimulate the conversion of 5' triphosphates to monophosphates by RppH. In addition, it suppresses the direct synthesis of monophosphorylated transcripts that begin with cytidine by reducing the cellular concentration of cytidine monophosphate, thereby disfavoring the 5'-terminal incorporation of this nucleotide by RNA polymerase during transcription initiation. Together, these findings suggest dual signaling pathways by which nucleotide metabolism can impact mRNA degradation in bacteria.
细菌细胞中 mRNA 降解的关键途径始于将初始 5' - 三磷酸转化为一磷酸,这种修饰使转录物更容易受到核糖核酸酶的攻击,这些核糖核酸酶对单磷酸化 5' 末端的亲和力增强了它们的催化效力。在大肠杆菌中,已知唯一对通过该途径控制降解重要的蛋白质是 RNA 焦磷酸水解酶 RppH、其异源二聚体伴侣 DapF 以及 5' - 单磷酸辅助内切酶 RNase E 和 RNase G。我们现在已经确定代谢酶胞苷酸激酶是另一种影响大肠杆菌中 5' - 端依赖的 mRNA 降解速率的蛋白质。它通过利用两种不同的机制来影响 RNA 的 5' - 末端磷酸化状态,每种机制都依赖于胞苷酸激酶的催化活性,而不仅仅是其在细胞中的存在。首先,该酶与 DapF 一起作用,通过 RppH 刺激 5' 三磷酸转化为一磷酸。此外,它通过降低细胞中胞苷一磷酸的浓度来抑制以胞苷起始的直接合成具有单磷酸化 5' 末端的转录物,从而在转录起始时不有利于 RNA 聚合酶将该核苷酸掺入 5' - 末端。这些发现共同表明核苷酸代谢可以通过两种信号通路影响细菌中的 mRNA 降解。