Key Laboratory of Medical Epigenetics and Metabolism, Fudan University Pudong Medical Center, Institutes of Biomedical Sciences, Fudan University , Shanghai, PR China.
Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, School of Basic Medical Sciences, Fudan University , Shanghai, PR China.
Virulence. 2020 Dec;11(1):1432-1442. doi: 10.1080/21505594.2020.1836910.
The proliferation of antibiotic resistance has its origins in horizontal gene transfer. The class 1 integrons mediate gene transfer by assimilating antibiotic-resistance genes through site-specific recombination. For the class 1 integrons the first assimilated gene normally encodes an aminoglycoside antibiotic resistance protein which is either an aminoglycoside acetyltransferase (AAC), nucleotidyltransferase - (ANT), or adenyl transferase (AAD). An aminoglycoside-sensing riboswitch RNA in the leader RNA of AAC/AAD that controls the expression of aminoglycoside resistance genes has been previously described. Here we explore the relationship between the recombinant products of integron recombination and a series of candidate riboswitch RNAs in the 5' UTR of (aminoglycoside adenyltransferases) genes. The RNA sequences from the 5' UTR of the genes from pathogenic strains that are the products of site-specific DNA recombination by class 1 integrons were investigated. Reporter assays, MicroScale Thermophoresis (MST) and covariance analysis revealed that a functional aminoglycoside-sensing riboswitch was selected at the DNA level through integron-mediated site-specific recombination. This study explains the close association between integron recombination and the aminoglycoside-sensing riboswitch RNA.
抗生素耐药性的扩散源于水平基因转移。类 1 整合子通过位点特异性重组同化抗生素耐药基因来介导基因转移。对于类 1 整合子,第一个被同化的基因通常编码一种氨基糖苷类抗生素耐药蛋白,该蛋白要么是氨基糖苷乙酰转移酶 (AAC)、核苷酸转移酶- (ANT),要么是腺嘌呤转移酶 (AAD)。先前已经描述了 AAC/AAD 启动子 RNA 中氨基糖苷感应核糖开关 RNA 控制氨基糖苷类耐药基因表达。在这里,我们探讨了整合子重组的重组产物与 5'UTR 中一系列候选核糖开关 RNA 之间的关系(氨基糖苷腺嘌呤转移酶)基因。研究了来自致病性菌株的 基因 5'UTR 的 RNA 序列,这些菌株是通过 1 类整合子的位点特异性 DNA 重组产生的。报告基因检测、微量热泳动 (MST) 和协方差分析表明,通过整合子介导的位点特异性重组,在 DNA 水平上选择了一种功能性氨基糖苷类感应核糖开关。这项研究解释了整合子重组与氨基糖苷感应核糖开关 RNA 之间的密切关联。