The Bifunctional Enzyme SpoT Is Involved in the Clarithromycin Tolerance of Helicobacter pylori by Upregulating the Transporters HP0939, HP1017, HP0497, and HP0471.
The Bifunctional Enzyme SpoT Is Involved in the Clarithromycin Tolerance of Helicobacter pylori by Upregulating the Transporters HP0939, HP1017, HP0497, and HP0471.
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双功能酶 SpoT 通过上调转运蛋白 HP0939、HP1017、HP0497 和 HP0471 参与幽门螺杆菌的克拉霉素耐受性
DOI：
10.1128/aac.02011-16
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发表时间：
2017-05
影响因子：
4.9
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Geng X;Li W;Chen Z;Gao S;Hong W;Ge X;Hou G;Hu Z;Zhou Y;Zeng B;Li W;Jia J;Sun Y
摘要克拉霉素(CLA)是根除幽门螺杆菌的常用药物。然而，对共轭亚油酸耐药的幽门螺杆菌的流行率正在上升。虽然23S rRNA的点突变是共轭亚油酸耐药的关键因素，但包括外排泵和调控基因在内的其他因素也参与了幽门螺杆菌对共轭亚油酸的耐药。幽门螺杆菌双功能酶斑点合成的鸟苷3‘-二磷酸-5’-三磷酸和鸟苷3‘，5’-二焦磷酸[(P)ppGpp]对一些细菌适应抗生素压力起着重要作用。然而，关于幽门螺杆菌的相关研究尚未见报道。此外，已发现转运蛋白与细菌耐药性有关。因此，本研究通过检测转运蛋白表达的变化来研究Spot在幽门螺杆菌对共轭亚油酸耐药中的作用，并探讨转运蛋白在幽门螺杆菌对共轭亚油酸耐药中的作用。本研究构建了一株ΔSPOT菌株，结果表明SPOT通过上调转运蛋白HP0939、HP1017、HP0497和HP0471参与了幽门螺杆菌的耐受。这是通过一系列分子和生化实验以及基因芯片进行评估的。此外，在抗性品系中，hp0939、hp0471和hp0497基因的敲除导致了对共轭亚油酸抗性的降低或丧失(后者在Δhp0497品系中)。此外，这四种转运蛋白在临床CLA耐药株中的平均表达水平显著高于临床CLA敏感株。综上所述，我们的结果揭示了幽门螺杆菌适应共轭亚油酸胁迫的一种新的分子机制。
ABSTRACT Clarithromycin (CLA) is a commonly recommended drug for Helicobacter pylori eradication. However, the prevalence of CLA-resistant H. pylori is increasing. Although point mutations in the 23S rRNA are key factors for CLA resistance, other factors, including efflux pumps and regulation genes, are also involved in the resistance of H. pylori to CLA. Guanosine 3′-diphosphate 5′-triphosphate and guanosine 3′,5′-bispyrophosphate [(p)ppGpp)], which are synthesized by the bifunctional enzyme SpoT in H. pylori, play an important role for some bacteria to adapt to antibiotic pressure. Nevertheless, no related research involving H. pylori has been reported. In addition, transporters have been found to be related to bacterial drug resistance. Therefore, this study investigated the function of SpoT in H. pylori resistance to CLA by examining the shifts in the expression of transporters and explored the role of transporters in the CLA resistance of H. pylori. A ΔspoT strain was constructed in this study, and it was shown that SpoT is involved in H. pylori tolerance of CLA by upregulating the transporters HP0939, HP1017, HP0497, and HP0471. This was assessed using a series of molecular and biochemical experiments and a cDNA microarray. Additionally, the knockout of genes hp0939, hp0471, and hp0497 in the resistant strains caused a reduction or loss (the latter in the Δhp0497 strain) of resistance to CLA. Furthermore, the average expression levels of these four transporters in clinical CLA-resistant strains were considerably higher than those in clinical CLA-sensitive strains. Taken together, our results revealed a novel molecular mechanism of H. pylori adaption to CLA stress.