RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究

doi:10.11843/j.issn.0366-6964.2023.09.019

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (9): 3793-3802.doi: 10.11843/j.issn.0366-6964.2023.09.019

RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究

李倩倩¹, 林馨¹, 韦依琳¹, 崔昊宇¹, 邹荣华², 吴晓妮², 葛家振², 黄国梁¹, 张丽娟¹, 郑福英², 蔺国珍^1*

1. 西北民族大学生命科学与工程学院, 兰州 730030;
2. 中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室, 兰州 730046

收稿日期:2022-10-26 发布日期:2023-09-22
通讯作者: 蔺国珍,主要从事细菌耐药机制、致病机制及诊断技术研究,E-mail:lgzh18@163.com
作者简介:李倩倩(1997-),女,陕西宝鸡人,硕士生,主要从事细菌耐药机制、致病机制及诊断技术研究,E-mail:y200130610@outlook.com;林馨(2003-),女,福建福州人,本科生,主要从事细菌耐药机制、致病机制及诊断技术研究,E-mail:2371796023@qq.com
基金资助:
国家自然科学基金(32260882);中央高校基本科研业务费专项资金(31920220137)

Study on the Role of RaeR in the Regulation of Efflux Pump RaeC-RaeA-RaeB of Rimerella anatipestifer

LI Qianqian¹, LIN Xin¹, WEI Yilin¹, CUI Haoyu¹, ZOU Ronghua², WU Xiaoni², GE Jiazhen², HUANG Guoliang¹, ZHANG Lijuan¹, ZHENG Fuying², LIN Guozhen^1*

1. College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China;
2. State Key Laboratory for Animal Disease Control and Prevention, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2022-10-26 Published:2023-09-22

摘要/Abstract

摘要： 旨在阐明鸭疫里默氏杆菌RA-LZ01株GE296_RS05175基因编码的RaeR对外排泵RaeC-RaeA-RaeB的调控作用。构建RA-LZ01株的GE296_RS05160基因(编码内膜蛋白RaeB)缺失株ΔRaeB和回复株cΔRaeB,以及GE296_RS05175基因缺失株ΔRaeR和回复株cΔRaeR。测定菌株的生长曲线、菌株对抗菌素的敏感性以及GE296_RS05160和GE296_RS05175基因在特异性底物刺激下的转录水平。结果显示,与亲本株相比,缺失株的生长能力无明显变化;与亲本株和回复株相比,缺失株ΔRaeB对卡那霉素、链霉素和十二烷基硫酸钠(sodium dodecyl sulfate,SDS)的敏感性上升,而缺失株ΔRaeR对卡那霉素、链霉素和SDS的敏感性降低;加入SDS刺激后,RA-LZ01和ΔRaeR株的GE296_RS05160基因的相对转录水平分别上升了2.26倍和4.64倍,ΔRaeR株中该基因的上调倍数明显超过了RA-LZ01株;在SDS刺激下,RA-LZ01和ΔRaeB株的GE296_RS05175基因的相对转录水平均明显下调。上述结果表明,外排泵RaeC-RaeA-RaeB与菌株的生长无明显相关性,可介导菌株对卡那霉素、链霉素和SDS产生耐受;底物SDS可刺激GE296_RS05175基因的转录水平降低,该基因编码的RaeR对RaeB的表达发挥负向调控作用。

关键词: 鸭疫里默氏杆菌, RaeB, RaeR, 调控

Abstract: The aim of this study was to clarify the role of RaeR encoded by GE296_RS05175 gene in the regulation of efflux pump RaeC-RaeA-RaeB of Rimerella anatipestifer RA-LZ01 strain. The GE296_RS05160 gene (encoding the inner membrane protein RaeB) deletion mutant ΔRaeB and complemented strain cΔRaeB of RA-LZ01 strain were constructed. Similarly, GE296_RS05175 gene deletion mutant ΔRaeR and its complemented strain cΔRaeR were constructed. The growth curves, antimicrobial susceptibility and transcription levels of GE296_RS05160 and GE296_RS05175 genes under specific substrate stimulation were measured. Compared with the parental strain, the growth abilities of the deletion mutants had no obvious change. By comparison with the parental strain and complemented strains, the sensitivity of ΔRaeB to kanamycin, streptomycin and (sodium dodecyl sulfate, SDS) were increased, while ΔRaeR showed the opposite results. After SDS stimulation, the relative transcription levels of GE296_RS05160 gene in RA-LZ01 and ΔRaeR strains increased 2.26 and 4.64 times, respectively. The upregulation of GE296_RS05160 gene in ΔRaeR strain was significantly higher than that in RA-LZ01 strain. Under SDS stimulation, the relative transcription levels of GE296_RS05175 gene in RA-LZ01 and ΔRaeB strains decreased significantly. These results indicated that the efflux pump RaeC-RaeA-RaeB had no significant correlation with the growth of RA-LZ01 strain, but could mediate the tolerance of the strain to kanamycin, streptomycin and SDS. The transcription level of GE296_RS05175 gene reduced under SDS pressure, and RaeR,which encoded by GE296_RS05175 gene, regulated the expression of RaeB negatively.

Key words: Rimerella anatipestifer, RaeB, RaeR, regulation

中图分类号:

S852.6

李倩倩, 林馨, 韦依琳, 崔昊宇, 邹荣华, 吴晓妮, 葛家振, 黄国梁, 张丽娟, 郑福英, 蔺国珍. RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究[J]. 畜牧兽医学报, 2023, 54(9): 3793-3802.

LI Qianqian, LIN Xin, WEI Yilin, CUI Haoyu, ZOU Ronghua, WU Xiaoni, GE Jiazhen, HUANG Guoliang, ZHANG Lijuan, ZHENG Fuying, LIN Guozhen. Study on the Role of RaeR in the Regulation of Efflux Pump RaeC-RaeA-RaeB of Rimerella anatipestifer[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3793-3802.

参考文献

[1] SEGERS P,MANNHEIM W,VANCANNEYT M,et al.Riemerella anatipestifer gen. nov., comb. nov., the causative agent of septicemia anserum exsudativa,and its phylogenetic affiliation within the Flavobacterium-Cytophaga rRNA homology group[J].Int J Syst Bacteriol,1993,43(4):768-776.
[2] MAGYAR T,GYURIS É,UJVÁRI B,et al.Genotyping of Riemerella anatipestifer by ERIC-PCR and correlation with serotypes[J].Avian Pathol,2019,48(1):12-16.
[3] UMAR Z,CHEN Q W,TANG B,et al.The poultry pathogen Riemerella anatipestifer appears as a reservoir for Tet(X) tigecycline resistance[J].Environ Microbiol,2021,23(12):7465-7482.
[4] 高海娇,程古月,王玉莲,等.细菌主要外排泵及其调控蛋白研究进展[J].畜牧兽医学报,2017,48(11):2023-2033.
GAO H J,CHENG G Y,WANG Y L,et al.Research progress of the mainly bacterial efflux pumps and related regulator[J].Acta Veterinaria et Zootechnica Sinica,2017,48(11):2023-2033.(in Chinese)
[5] LI X Z,PLÉSIAT P,NIKAIDO H.The challenge of efflux-mediated antibiotic resistance in Gram-negative bacteria[J].Clin Microbiol Rev,2015,28(2):337-418.
[6] PUZARI M,CHETIA P.RND efflux pump mediated antibiotic resistance in Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa:a major issue worldwide[J].World J Microbiol Biotechnol,2017,33(2):24.
[7] ZHANG X,WANG M S,LIU M F,et al.Contribution of RaeB,a putative RND-type transporter to aminoglycoside and detergent resistance in Riemerella anatipestifer[J].Front Microbiol,2017,8:2435.
[8] WANG Y P,LI S D,GONG X W,et al.Characterization of RaeE-RaeF-RopN,a putative RND efflux pump system in Riemerella anatipestifer[J].Vet Microbiol,2020,251:108852.
[9] 秦明星,宫晓炜,陈启伟,等.鸭疫里默氏杆菌RA-LZ01株GE296_RS01450基因介导其对重金属离子产生耐受性的分析[J].畜牧兽医学报,2021,52(10):2864-2873.
QIN M X,GONG X W,CHEN Q W,et al.Study on the GE296_RS01450 gene-mediated tolerance of Rimerella anatipestifer strain RA-LZ01 to heavy metal ions[J].Acta Veterinaria et Zootechnica Sinica,2021,52(10):2864-2873.(in Chinese)
[10] 吉果,陈启伟,宫晓炜,等.常用抗生素对鸭疫里默氏杆菌MIC最佳试验条件的确立[J].黑龙江畜牧兽医,2017(3):32-36,40.
JI G,CHEN Q W,GONG X W,et al.Determination of the optimal testing conditions for the MICs of common antibiotics against Riemerella anatipestifer[J].Heilongjiang Animal Science and Veterinary Medicine,2017(3):32-36,40.(in Chinese)
[11] GRIMSEY E M,FAIS C,MARSHALL R L,et al.Chlorpromazine and amitriptyline are substrates and inhibitors of the AcrB multidrug efflux pump[J].mBio,2020,11(3):e00465-20.
[12] AIRES J R,NIKAIDO H.Aminoglycosides are captured from both periplasm and cytoplasm by the AcrD multidrug efflux transporter of Escherichia coli[J].J Bacteriol,2005,187(6):1923-1929.
[13] LAU S Y,ZGURSKAYA H I.Cell division defects in Escherichia coli deficient in the multidrug efflux transporter AcrEF-TolC[J].J Bacteriol,2005,187(22):7815-7825.
[14] MOSENG M A,LYU M,PIPATPOLKAI T,et al.Cryo-EM structures of CusA reveal a mechanism of metal-ion export[J].mBio,2021,12(2):e00452-21.
[15] XUAN W K,RODRÍGUEZ-BLANCO G,SOUTHAM A D,et al.Metabolomics reveal potential natural substrates of AcrB in Escherichia coli and Salmonella enterica Serovar Typhimurium[J].mBio,2021,12(2):e00109-21.
[16] CUESTA BERNAL J,EL-DELIK J,GÖTTIG S,et al.Characterization and molecular determinants for β-Lactam specificity of the multidrug efflux pump AcrD from Salmonella typhimurium[J].Antibiotics (Basel),2021,10(12):1494.
[17] EAVES D J,RICCI V,PIDDOCK L J V.Expression of acrB,acrF,acrD,marA,and soxS in Salmonella enterica serovar typhimurium:role in multiple antibiotic resistance[J].Antimicrob Agents Chemother,2004,48(4):1145-1150.
[18] PESINGI P V,SINGH B R,PESINGI P K,et al.MexAB-OprM efflux pump of Pseudomonas aeruginosa offers resistance to carvacrol:a herbal antimicrobial agent[J].Front Microbiol,2019,10:2664.
[19] JEANNOT K,ELSEN S,KÖHLER T,et al.Resistance and virulence of Pseudomonas aeruginosa clinical strains overproducing the MexCD-OprJ efflux pump[J].Antimicrob Agents Chemother,2008,52(7):2455-2462.
[20] VAILLANCOURT M,LIMSUWANNAROT S P,BRESEE C,et al.Pseudomonas aeruginosa mexR and mexEF antibiotic efflux pump variants exhibit increased virulence[J].Antibiotics (Basel),2021,10(10):1164.
[21] DEY D,KAVANAUGH L G,CONN G L.Antibiotic substrate selectivity of Pseudomonas aeruginosa MexY and MexB efflux systems is determined by a goldilocks affinity[J].Antimicrob Agents Chemother,2020,64(8):e00496-20.
[22] WESTON N,SHARMA P,RICCI V,et al.Regulation of the AcrAB-TolC efflux pump in Enterobacteriaceae[J].Res Microbiol,2018,169(7-8):425-431.
[23] HIRAKAWA H,INAZUMI Y,MASAKI T,et al.Indole induces the expression of multidrug exporter genes in Escherichia coli[J].Mol Microbiol,2005,55(4):1113-1126.
[24] NISHINO K,YAMAGUCHI A.Role of histone-like protein H-NS in multidrug resistance of Escherichia coli[J].J Bacteriol,2004,186(5):1423-1429.
[25] NISHINO K,HAYASHI-NISHINO M,YAMAGUCHI A.H-NS modulates multidrug resistance of Salmonella enterica serovar Typhimurium by repressing multidrug efflux genes acrEF[J].Antimicrob Agents Chemother,2009,53(8):3541-3543.
[26] OLLIVER A,VALLÉ M,CHASLUS-DANCLA E,et al.Overexpression of the multidrug efflux operon acrEF by insertional activation with IS1 or IS10 elements in Salmonella enterica serovar typhimurium DT204 acrB mutants selected with fluoroquinolones[J].Antimicrob Agents Chemother,2005,49(1):289-301.

RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究

Study on the Role of RaeR in the Regulation of Efflux Pump RaeC-RaeA-RaeB of Rimerella anatipestifer

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	郭妍婷, 周建民, 王晶, 齐广海. 产蛋鸡子宫部钙离子转运及调控的研究进展[J]. 畜牧兽医学报, 2024, 55(1): 39-50.
[2]	王栋梁, 任静, 郝琴琴, 李鹏飞. 牛CART基因核心启动子鉴定及转录调控分析[J]. 畜牧兽医学报, 2023, 54(9): 3689-3699.
[3]	吴志立, 姚军虎, 雷新建. 过瘤胃葡萄糖对围产期奶畜营养调控的研究进展[J]. 畜牧兽医学报, 2023, 54(8): 3173-3182.
[4]	王思盈, 邹宏, 宋振辉. Na⁺/H⁺交换体家族第三个亚型在感染性腹泻中的作用及活性调控机制[J]. 畜牧兽医学报, 2023, 54(8): 3230-3241.
[5]	郭艳丽, 李可强, 白少川, 王涛, 王德贺, 王麒, 李兰会. ALV-E的结构、活性调控以及对宿主功能的影响[J]. 畜牧兽医学报, 2023, 54(7): 2683-2691.
[6]	禹世雄, 魏凌云, 徐甜甜, 焦金真, 蒋林树, 贺志雄. 幼龄反刍动物肠道微生物定植规律及其营养调控研究进展[J]. 畜牧兽医学报, 2023, 54(7): 2701-2707.
[7]	安宗麒, 占思远, 李利, 张红平. circRNA作为ceRNA调控畜禽重要经济性状的研究进展[J]. 畜牧兽医学报, 2023, 54(6): 2215-2222.
[8]	陈松彪, 尚珂, 杜付熙, 余祖华, 李静, 贾艳艳, 廖成水, 张春杰, 丁轲, 程相朝. 沙门菌VI型分泌系统组装、结构特征和分泌调控网络的研究进展[J]. 畜牧兽医学报, 2023, 54(6): 2252-2263.
[9]	王唯, 贺小云, 储明星. 昼夜节律与雌激素协同调控哺乳动物生殖的研究进展[J]. 畜牧兽医学报, 2023, 54(5): 1771-1781.
[10]	常心雨, 王继光, 王晶, 张海军, 齐广海, 邱凯, 武书庚. 商品蛋鸡精准饲养技术研究进展[J]. 畜牧兽医学报, 2023, 54(5): 1815-1823.
[11]	龙琴琴, 魏敏, 王雨婷, 文明, 庞峰. 羊口疮病毒与宿主的博弈:免疫应答与病毒免疫逃逸机制[J]. 畜牧兽医学报, 2023, 54(5): 1845-1853.
[12]	王岚, 何明宇, 张敏, 丁军涛. MicroRNA调控抗病毒免疫和病毒复制[J]. 畜牧兽医学报, 2023, 54(2): 463-472.
[13]	卓儒浩, 柳清扬, 钟翔. 姜黄素调控肠道菌群及抗病毒作用研究进展[J]. 畜牧兽医学报, 2023, 54(2): 473-483.
[14]	邢文文, 齐南南, 李梦轩, 刘吉英. YY1作用机制及在动物繁殖调控中的研究进展[J]. 畜牧兽医学报, 2023, 54(10): 4040-4049.
[15]	戴超辉, 李辉, 赵为民, 付言峰, 廖超, 李碧侠, 王学敏, 程金花. 脱氧雪腐镰刀菌烯醇对猪毒害作用及毒理机制研究进展[J]. 畜牧兽医学报, 2023, 54(1): 24-35.