title: The Bifunctional Enzyme SpoT Is Involved in the Clarithromycin Tolerance of Helicobacter pylori by Upregulating the Transporters HP0939, HP1017, HP0497, and HP0471.
abstract: Clarithromycin (CLA) is a commonly recommended drug forHelicobacter pylorieradication. However, the prevalence of CLA-resistantH. pyloriis 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 ofH. pylorito CLA. Guanosine 3'-diphosphate 5'-triphosphate and guanosine 3',5'-bispyrophosphate [(p)ppGpp)], which are synthesized by the bifunctional enzyme SpoT inH. pylori, play an important role for some bacteria to adapt to antibiotic pressure. Nevertheless, no related research involvingH. pylorihas been reported. In addition, transporters have been found to be related to bacterial drug resistance. Therefore, this study investigated the function of SpoT inH. pyloriresistance to CLA by examining the shifts in the expression of transporters and explored the role of transporters in the CLA resistance ofH. pyloriA ΔspoTstrain was constructed in this study, and it was shown that SpoT is involved inH. pyloritolerance 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 geneshp0939,hp0471, andhp0497in the resistant strains caused a reduction or loss (the latter in the Δhp0497strain) 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 ofH. pyloriadaption to CLA stress.
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Xiwen
authors.initials: X
authors.lastname: Geng
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Wen
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Microbiology, Guiyang Medical University, Guiyang, Guizhou, People's Republic of China.
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Sizhe
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Microbiology, Guiyang Medical University, Guiyang, Guizhou, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hong
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Xiaoran
authors.initials: X
authors.lastname: Ge
authors.affiliation: Laboratory of Experimental Teratology, Ministry of Education and Institute of Experimental Nuclear Medicine, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Guihua
authors.initials: G
authors.lastname: Hou
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Zhekai
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Yabin
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Beini
authors.initials: B
authors.lastname: Zeng
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China.
authors.forename: Jihui
authors.initials: J
authors.lastname: Jia
authors.affiliation: Department of Microbiology, School of Medicine, Shandong University, Jinan, Shandong, People's Republic of China syd@sdu.edu.cn.
authors.forename: Yundong
authors.initials: Y
authors.lastname: Sun
mesh_terms: D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D017291:Clarithromycin / Q000627:therapeutic use*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006158:Guanosine Pentaphosphate / Q000378:metabolism; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D016481:Helicobacter Infections / Q000188:drug therapy* / Q000382:microbiology; D016480:Helicobacter pylori / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D064251:Multifunctional Enzymes / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D064251:Multifunctional Enzymes; D006159:Guanosine Tetraphosphate; D006158:Guanosine Pentaphosphate; D017291:Clarithromycin
keywords: Helicobacter pylori; SpoT; antibiotic resistance; clarithromycin; transporters