Role of AlgC and GalU in the Intrinsic Antibiotic Resistance of Helicobacter pylori

Abstract

Shunhang Feng,1,2,* Jiansheng Lin,1,2,* Xiaoyan Zhang,1,2 Xin Hong,1,2 Wanyin Xu,1,2 Yancheng Wen,1,2 Feifei She1,2 1Key Laboratory of Gastrointestinal Cancer (Fujian Medical University), Ministry of Education, Fuzhou, People’s Republic of China; 2Fujian Key Laboratory of Tumor Microbiology, Department of Medical Microbiology, School for Basic Medical Sciences, Fujian Medical University, Fuzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Feifei She, Fujian Medical University, Xueyuan Road 1, Minhou County, Fuzhou, Fujian Province, People’s Republic of China, Tel [phone], Email [email protected] Yancheng Wen, Fujian Medical University, Xueyuan Road 1, Minhou County, Fuzhou, Fujian Province, People’s Republic of China, Tel [phone], Email [email protected]: Helicobacter pylori is associated with the development of gastrointestinal diseases. However, its eradication is challenged by an increased rate of drug resistance. AlgC and GalU are important for the synthesis of UDP-glucose, which is a substrate for the synthesis of lipopolysaccharide (LPS) in H. pylori. In this study, we investigated the role of UDP-glucose in the intrinsic drug resistance in H. pylori.Methods: Gene knockout strains or complementation strains, including ΔalgC, ΔgalU, ΔgalE, Δhp0045, ΔalgC/algC* and ΔgalU/galU* were constructed in Hp26695; and ΔalgC and ΔgalU were also constructed in two clinical drug-resistant strains, Hp008 and Hp135. The minimum inhibitory concentrations (MIC) of H. pylori to amoxicillin (AMO), tetracycline (TET), clarithromycin (CLA), metronidazole (MNZ), levofloxacin (LEV), and rifampicin (RIF) were measured using MIC Test Strips. Silver staining was performed to examine the role of AlgC and GalU in LPS synthesis. Ethidium bromide (EB) accumulation assay was performed to assess the outer membrane permeability of H. pylori strains.Results: Knockout of algC and galU in H. pylori resulted in increased drug sensitivity to AMO, MNZ, CLA, LEV, and RIF; whereas knockout of hp0045 and galE, which are involved in GDP-fucose and UDP-galactose synthesis, respectively, did not significantly alter the drug sensitivity of H. pylori. Knockout of algC and galU in clinically drug-resistant strains resulted in significantly increased drug sensitivity to all the antibiotics, except MNZ. The lipid A-core structure was altered in ΔalgC and ΔgalU when their EB accumulation was higher than that in the wild type and complementation strains.Conclusion: UDP-glucose may play an important role in increasing drug resistance to AMO, MNZ, CLA, LEV, TET, and RIF by maintaining the lipid A-core structure and decreasing membrane permeability. AlgC and GalU may serve as potential drug targets for decreasing antibiotic resistance in clinical isolates.Keywords: H. pylori, UDP-glucose, algC, galU, intrinsic multidrug resistance

Keywords

• h. pylori
• udp-glucose
• algc
• galu
• intrinsic multidrug resistance