Radical SAM-Mediated Methylation of Ribosomal RNA.
Radical SAM-Mediated Methylation of Ribosomal RNA.
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DOI：
10.1016/bs.mie.2015.03.002
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发表时间：
2015
影响因子：
--
通讯作者：
Fujimori DG
中科院分区：
文献类型：
--
作者：
Stojković V;Fujimori DG
RNA 的转录后修饰在广泛的生物过程中发挥着重要作用。在核糖体 RNA (rRNA) 中，核苷酸碱基的甲基化是主要的修饰。近年来，细菌 23S rRNA 中腺苷 2503 (A2503) 的甲基化引起了人们的广泛关注，这是由于甲基掺入的不寻常的区域选择性以及由此产生的甲基化的病理生理作用。具体来说，A2503 在腺嘌呤环的 C2 和 C8 位点被甲基化，引入的修饰分别对翻译保真度和抗生素耐药性产生深远影响。这些修饰是由最近发现的自由基 S-腺苷甲硫氨酸（自由基 SAM）甲基合酶类的两个成员 RlmN 和 Cfr 进行的。在这里，我们提出了几种可用于评估这些酶在体内和体外条件下的活性的方法。
Post-transcriptional modifications of RNA play an important role in a wide range of biological processes. In ribosomal RNA (rRNA), methylation of nucleotide bases is the predominant modification. In recent years, methylation of adenosine 2503 (A2503) in bacterial 23S rRNA has attracted significant attention due to both the unusual regioselectivity of the methyl group incorporation, as well as the pathophysiological roles of the resultant methylations. Specifically, A2503 is methylated at the C2 and C8 positions of the adenine ring, and the introduced modifications have a profound impact on translational fidelity and antibiotic resistance, respectively. These modifications are performed by RlmN and Cfr, two members, of the recently discovered class of radical S-adenosylmethionine (radical SAM) methylsynthases. Here, we present several methods that can be used to evaluate the activity of these enzymes, under both in vivo and in vitro conditions.