Antibiotic resistance evolved via inactivation of a ribosomal RNA methylating enzyme.
Antibiotic resistance evolved via inactivation of a ribosomal RNA methylating enzyme.
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DOI：
10.1093/nar/gkw699
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发表时间：
2016-10-14
影响因子：
14.9
通讯作者：
Fujimori DG
中科院分区：
文献类型：
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作者：
Stojković V;Noda-Garcia L;Tawfik DS;Fujimori DG
细菌核糖体的修饰调节核糖体的功能并调节其对抗生素的敏感性。通过修饰23S rRNA中高度保守的腺苷A2503，甲基化酶Cfr赋予对一系列核糖体靶向抗生素的抗性。相同的腺苷也被RlmN甲基化，RlmN是一种广泛分布于细菌中的酶。当RlmN修饰C2时，Cfr修饰A2503的C8位置。RlmN和Cfr之间共享的核苷酸底物和系统发育关系促使我们研究该酶家族中抗生素抗性的进化起源。使用抗生素选择下的RlmN的定向进化，我们获得了介导低水平抗性的RlmN变体。令人惊讶的是，这些变体不是通过Cfr样C8甲基化，而是通过抑制A2503的内源性RlmN C2甲基化来赋予抗性。在临床耐药分离株中检测到RlmN失活突变表明，体外进化变体使用的机制在临床环境中也是相关的。此外，如系统发育分析所示，似乎Cfr并不偏离RlmN家族，而是来自另一个不同的预测自由基SAM甲基化酶家族，其功能仍然未知。
Modifications of the bacterial ribosome regulate the function of the ribosome and modulate its susceptibility to antibiotics. By modifying a highly conserved adenosine A2503 in 23S rRNA, methylating enzyme Cfr confers resistance to a range of ribosome-targeting antibiotics. The same adenosine is also methylated by RlmN, an enzyme widely distributed among bacteria. While RlmN modifies C2, Cfr modifies the C8 position of A2503. Shared nucleotide substrate and phylogenetic relationship between RlmN and Cfr prompted us to investigate evolutionary origin of antibiotic resistance in this enzyme family. Using directed evolution of RlmN under antibiotic selection, we obtained RlmN variants that mediate low-level resistance. Surprisingly, these variants confer resistance not through the Cfr-like C8 methylation, but via inhibition of the endogenous RlmN C2 methylation of A2503. Detection of RlmN inactivating mutations in clinical resistance isolates suggests that the mechanism used by the in vitro evolved variants is also relevant in a clinical setting. Additionally, as indicated by a phylogenetic analysis, it appears that Cfr did not diverge from the RlmN family but from another distinct family of predicted radical SAM methylating enzymes whose function remains unknown.
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自由基 SAM 酶进行甲基转移的结构基础。
DOI：
10.1126/science.1205358
发表时间：
2011-05-27
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Boal AK;Grove TL;McLaughlin MI;Yennawar NH;Booker SJ;Rosenzweig AC
通讯作者：
Rosenzweig AC
DOI：
10.1016/j.ijantimicag.2005.11.008
发表时间：
2006-04-01
影响因子：
10.8
作者：
Gales, Ana C.;Sader, Helio S.;Barth, Afonso L.
通讯作者：
Barth, Afonso L.
影响因子：
4
作者：
McCusker, Kevin P.;Fujimori, Danica Galonic
通讯作者：
Fujimori, Danica Galonic
影响因子：
14.9
作者：
Kaminska KH;Purta E;Hansen LH;Bujnicki JM;Vester B;Long KS
通讯作者：
Long KS
大肠杆菌K-12的构造框架，单基因敲除突变体：Keio Collection。
影响因子：
9.9
作者：
通讯作者：
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