Pseudouridimycin (PUM) is a selective nucleoside-analog inhibitor of bacterial RNA polymerase with activity against Gram-positive and Gram-negative bacteria. PUM, produced byStreptomycessp. ID38640, consists of a formamidinylated,N-hydroxylated Gly-Gln dipeptide conjugated to 5′-aminopseudouridine. We report the characterization of the PUM gene cluster. Bioinformatic analysis and mutational knockouts ofpumgenes with analysis of accumulated intermediates, define the PUM biosynthetic pathway. The work provides the first biosynthetic pathway of aC-nucleoside antibiotic and reveals three unexpected features: production of free pseudouridine by the dedicated pseudouridine synthase, PumJ; nucleoside activation by specialized oxidoreductases and aminotransferases; and peptide-bond formation by amide ligases. A central role in the PUM biosynthetic pathway is played by the PumJ, which represents a divergent branch within the TruD family of pseudouridine synthases. PumJ-like sequences are associated with diverse gene clusters likely to govern the biosynthesis of different classes ofC-nucleoside antibiotics.

中文翻译：

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伪尿嘧啶生物合成途径的分析提供了对C-核苷抗生素形成的见解

伪尿di霉素（PUM）是细菌RNA聚合酶的选择性核苷类似物抑制剂，对革兰氏阳性和革兰氏阴性细菌具有活性。PUM，由Streptomycessp生产。ID38640，由与5'-氨基伪尿苷缀合的甲酰胺基化，N-羟基化的Gly-Gln二肽组成。我们报告了P​​UM基因簇的表征。生物信息学分析和对pumgenes的突变敲除以及对积累的中间体的分析，定义了PUM生物合成途径。这项工作提供了αC-核苷抗生素的第一个生物合成途径，并揭示了三个出乎意料的特征：专用假尿苷合酶PumJ产生游离的假尿苷；通过专门的氧化还原酶和氨基转移酶激活核苷；酰胺连接酶形成肽键。PumJ在PUM生物合成途径中起着核心作用，PumJ代表拟尿苷合酶的TruD家族内的一个分歧分支。PumJ样序列与可能控制不同类别​​的C-核苷抗生素的生物合成的各种基因簇相关。