鼠伤寒沙门菌BaeSR对氟喹诺酮类药物耐药性的调控机制

doi:10.11843/j.issn.0366-6964.2022.03.022

Abstract

Abstract: BaeSR is an important two component system in Salmonella Typhimurium, which affects bacterial resistance by regulating the transcription and expression of downstream target genes.The aim of this study was to explore the regulatory mechanism of BaeR overexpression in S. Typhimurium to fluoroquinolones (FQs), and to find the potential target gene of BaeR. In this study, we first expressed, purified and validated the BaeR protein, and screened the drug-related differentially expressed genes (include ompW, sodB, tolC, mdtD, spy and marR) from transcriptomics as potential target genes of BaeR. Combined with LacZ reporter gene fusion assay and electrophoretic mobility shift assay (EMSA), we studied the resistance mechanism of BaeSR to S. Typhimurium. The target band appeared at 28 ku after purification, which indicated that BaeR protein was successfully expressed. LacZ reporter gene fusion assay results show that, BaeR can positively regulate the expression of ompW, sodB, tolC, mdtD, spy and marR after overexpression. EMSA results indicate that BaeR protein can directly bind to the promoter region of marR and regulate the expression of marR gene. In conclusion, BaeSR can positively regulate the transcriptional expression of target genes ompW, sodB, tolC, mdtD, spy and marR in S. Typhimurium, and directly regulate the expression of marR, thus affecting the resistance of S. Typhimurium to FQs.

Key words: Salmonella Typhimurium, BaeSR, drug resistance, EMSA

CLC Number:

S852.61

ZHANG Lin, LU Fang, FU Hengfeng, JIANG Xidi, WEI Qiling, QI Caili, GAO Haixia, LI Lin. Regulation Mechanism of BaeSR on Fluoroquinolones Resistance in Salmonella Typhimurium[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 894-903.

References 32

[1]	吕强华.丁香醛对沙门氏菌Ⅲ型分泌系统的抑制作用及机制[D].长春:吉林大学, 2020.LV Q H. Inhibitory effect and mechanism of syringaldehyde against Salmonella type Ⅲ secretion system[D]. Changchun:Jilin University, 2020.(in Chinese)
[2]	程春荣,安戈,罗琦,等. 145株沙门氏菌耐药性分析[J].河南预防医学杂志, 2012, 23(4):256-258, 261.CHENG C R, AN G, LUO Q, et al. Analysis of antimicrobial susceptibility of 145 Salmonella isolates[J]. Henan Journal of Preventive Medicine, 2012, 23(4):256-258, 261.(in Chinese)
[3]	XIA S L, HENDRIKSEN R S, XIE Z Q, et al. Molecular characterization and antimicrobial susceptibility of Salmonella isolates from infections in humans in Henan Province, China[J]. J Clin Microbiol, 2009, 47(2):401-409.
[4]	FERRARI R G, ROSARIO D K A, CUNHA-NETO A, et al. Worldwide epidemiology of Salmonella serovars in animal-based foods:a meta-analysis[J]. Appl Environ Microbiol, 2019, 85(14):e00591-19.
[5]	CASTRO-VARGAS R E, HERRERA-SÁNCHEZ M P, RODRÍGUEZ-HERNÁNDEZ R, et al. Antibiotic resistance in Salmonella spp. isolated from poultry:a global overview[J]. Vet World, 2020, 13(10):2070-2084.
[6]	李琳,沈建忠,吴永宁,等.耐环丙沙星沙门菌marA、soxS和ramA基因以及外排泵基因acrA和acrB的表达水平分析[J].中国兽医科学, 2009, 39(10):928-932.LI L, SHEN J Z, WU Y N, et al. Expression of marA, soxS and ramA genes and efflux pump genes acrA and acrB in ciprofloxacin-resistant Salmonella[J]. Chinese Veterinary Science, 2009, 39(10):928-932.(in Chinese)
[7]	孟娇,陈晶瑜. Rcs双组分调节系统对细菌环境应答的分子调控研究进展[J].微生物学通报, 2021, 48(1):210-222.MENG J, CHEN J Y. Role of Rcs two-component regulatory system in environmental responses of bacteria:a review[J]. Microbiology China, 2021, 48(1):210-222.(in Chinese)
[8]	STOCK A M, ROBINSON V L, GOUDREAU P N. Two-component signal transduction[J]. Annu Rev Biochem, 2000, 69:183-215.
[9]	BARANOVA N, NIKAIDO H. The baeSR two-component regulatory system activates transcription of the yegMNOB (mdtABCD) transporter gene cluster in Escherichia coli and increases its resistance to novobiocin and deoxycholate[J]. J Bacteriol, 2002, 184(15):4168-4176.
[10]	NISHINO K, NIKAIDO E, YAMAGUCHI A. Regulation of multidrug efflux systems involved in multidrug and metal resistance of Salmonella enterica serovar Typhimurium[J]. J Bacteriol, 2007, 189(24):9066-9075.
[11]	WANG S Y, YOU C B, MEMON F Q, et al. BaeR participates in cephalosporins susceptibility by regulating the expression level of outer membrane proteins in Escherichia coli[J]. J Biochem, 2021, 169(1):101-108.
[12]	ISLAM S, ZHOU Y, LIANG L, et al. Application of a phage cocktail for control of Salmonella in foods and reducing biofilms[J]. Viruses, 2019, 11(9):841.
[13]	RUGBJERG H, WINGSTRAND A, HALD T, et al. Estimating the number of undetected multi-resistant Salmonella Typhimurium DT104 infected pig herds in Denmark[J]. Prev Vet Med, 2004, 65(3-4):147-171.
[14]	贾瑞瑞. Dip2a-LacZ报告基因小鼠模型的制作及表达分析[D].长春:东北师范大学, 2015.JIA R R. Generation of Dip2a-LacZ reporter mouse model and analyzing Dip2a expression pattern[D]. Changchun:Northeast Normal University, 2015.(in Chinese)
[15]	HAMED A A, KHEDR M, ABDELRAOF M. Activation of LacZ gene in Escherichia coli DH5α viaα-complementation mechanism for β-galactosidase production and its biochemical characterizations[J]. J Genet Eng Biotechnol, 2020, 18(1):80.
[16]	付立霞,徐敬潇,韩先干,等.一种基于lacZ基因和pUC复制子的原核启动子报告系统的构建及评价[J].生物工程学报, 2021, 37(1):321-330.FU L X, XU J X, HAN X G, et al. Construction and evaluation of a pUC-type prokaryotic promoter reporter system based on lacZ gene[J]. Chinese Journal of Biotechnology, 2021, 37(1):321-330.(in Chinese)
[17]	AVIV G, GAL-MOR O. lacZ reporter system as a tool to study virulence gene regulation in bacterial pathogens[J]. Methods Mol Biol, 2018, 1734:39-45.
[18]	JUERS D H, MATTHEWS B W, HUBER R E. LacZ β-galactosidase:structure and function of an enzyme of historical and molecular biological importance[J]. Protein Sci, 2012, 21(12):1792-1807.
[19]	王慧琦.野油菜黄单胞菌 β半乳糖苷酶缺失菌株的构建及双组分系统RavA/RavR调控鞭毛合成通路的初步研究[D].海口:海南大学, 2019.WANG H Q. Construction of a galactosidase-deficient strain of Xanthomonas campestris and preliminary study on the regulation of flagellar synthesis by the two-component system RavA/RavR[D]. Haikou:Hainan University, 2019.(in Chinese)
[20]	仇越,杜天钰,王迎,等.副溶血弧菌密度感应系统调控exsA和exsD的分子机制[J].军事医学, 2020, 44(10):740-745.QIU Y, DU T Y, WANG Y, et al. Regulation of exsA and exsD by quorum sensing in Vibrio parahaemolyticus[J]. Military Medical Sciences, 2020, 44(10):740-745.(in Chinese)
[21]	陈冠霖,王怡旻,范静,等. RNA结合蛋白研究技术[J].生命科学, 2021, 33(3):303-311.CHEN G L, WANG Y M, FAN J, et al. Technologies of studying RNA binding proteins[J]. Chinese Bulletin of Life Sciences, 2021, 33(3):303-311.(in Chinese)
[22]	陈传荣.鼠伤寒沙门氏菌HilD对ssrAB的调控作用[D].合肥:安徽农业大学, 2017.CHEN C R. Regulation of Salmonella typhimurium HilD on ssrAB[D]. Hefei:Anhui Agricultural University, 2017.(in Chinese)
[23]	YU L M, LI W C, XUE M, et al. Regulatory role of the two-component system BasSR in the expression of the EmrD multidrug efflux in Escherichia coli[J]. Microb Drug Resist, 2020, 26(10):1163-1173.
[24]	YU L M, LI W C, LIU Z C, et al. Role of McbR in the regulation of antibiotic susceptibility in avian pathogenic Escherichia coli[J]. Poult Sci, 2020, 99(12):6390-6401.
[25]	NISHINO K, HONDA T, YAMAGUCHI A. Genome-wide analyses of Escherichia coli gene expression responsive to the BaeSR two-component regulatory system[J]. J Bacteriol, 2005, 187(5):1763-1772.
[26]	YE Y W, LING N, GAO J N, et al. Roles of outer membrane protein W (OmpW) on survival, morphology, and biofilm formation under NaCl stresses in Cronobacter sakazakii[J]. J Dairy Sci, 2018, 101(5):3844-3850.
[27]	陈妍妍.溶藻弧菌定植因子AcfA的功能及其对SOD、Flg和Dct的调控[D].湛江:广东海洋大学, 2019.CHEN Y Y. Study on the function and regulation of accessory colonization factor AcfA of Vibrio alginolyticus to SOD, Flg and Dct[D]. Zhanjiang:Guangdong Ocean University, 2019.(in Chinese)
[28]	LEBLANC S K D, OATES C W, RAIVIO T L. Characterization of the induction and cellular role of the BaeSR two-component envelope stress response of Escherichia coli[J]. J Bacteriol, 2011, 193(13):3367-3375.
[29]	ROSNER J L, MARTIN R G. Reduction of cellular stress by TolC-dependent efflux pumps in Escherichia coli indicated by BaeSR and CpxARP activation of spy in efflux mutants[J]. J Bacteriol, 2013, 195(5):1042-1050.
[30]	张传珍,常满霞,杨磊,等. MarA和SoxS共同调节大肠杆菌K12外排泵AcrAB-TolC的表达[J].中国兽医学报, 2017, 37(12):2370-2377.ZHANG C Z, CHANG M X, YANG L, et al. Co-regulation of MarA and SoxS on expression of AcrAB-TolC in development of ciprofloxacin resistance in E. coli K12[J]. Chinese Journal of Veterinary Science, 2017, 37(12):2370-2377.(in Chinese)
[31]	GRKOVIC S, BROWN M H, SKURRAY R A. Regulation of bacterial drug export systems[J]. Microbiol Mol Biol Rev, 2002, 66(4):671-701.
[32]	李琳,王玉平,吴永宁,等.沙门氏菌环丙沙星诱导株marR, soxR和acrR基因突变与多重耐药性的相关性研究(英文)[J].食品安全质量检测技术, 2009, 1(1):1-9.LI L, WANG Y P, WU Y N, et al. Association of marR, soxR and acrR mutations with multiple antibiotic resistance in ciprofloxacin-selected Salmonella mutants[J]. Food Safety and Quality Detection Technology, 2009, 1(1):1-9.

Regulation Mechanism of BaeSR on Fluoroquinolones Resistance in Salmonella Typhimurium

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 32

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHENG Rui, LIU Zishi, ZHANG Kangyou, YAN Yong, WEI Ling, ZEREN Wengmu, DINGZE Demi, HUANG Jianjun, WANG Li, WEI Yong. Isolation, Identification and Biological Characterization of Colletotrichum jasminigenum in Stems of Peanuts [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2206-2213.
[2]	ZHANG Yanmin, ZHAO Dongxu, WANG Wenlong. Mechanism of Resistance to Ivermectin in the Haemonchus contortus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1511-1520.
[3]	WANG Dong, LIU Kexin, HE Yanjun, DENG Shouxiang, LIU Yun, MA Weiming. Effects of Dietary Sodium Humate Supplementation on Liver Tissue Inflammation and Antioxidant Capacity of Salmonella Typhimurium-Infected Broilers [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 629-639.
[4]	LI Zhaolong, KONG Xiangrui, LIN Fengqiang, WANG Xiuping, ZHAO Ran, PENG Xiaoli, CHEN Changsong. Preliminary Study of the Mechanism of Inhibition of Salmonella Typhimurium by Ec-12 Supernatant [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 739-750.
[5]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[6]	ZHAO Feifei, LI Jie, HAN Ning, XIE Shiting, ZENG Zhenling. Antibacterial Drug Resistance Analysis of Klebsiella pneumoniae Isolated from Slaughterhouse [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3044-3053.
[7]	QIN Lei, WU Huimin, XU Qiqi, CHEN Wanzhao, WANG Dong, LI Hongbo, XIA Panpan, LIU Zepeng, XIA Lining. Effect of Exogenous Drug-Resistant Salmonella Typhimurium on Intestinal Flora in Healthy Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2158-2169.
[8]	JIANG Zenghai, TENG Lin, HE Anwen, LIU Yanyan, YUE Min, HE Qigai. Genomic Analysis of Salmonella Typhimurium Isolates and Salmonella Serotype 4, [5], 12: i:- Isolates from Pig-borne Food Chain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1199-1209.
[9]	Lü Ruoyi, SI Xiaohui, SUN Zhigang, SHI Xiaomin, LIU Xiaoye. Drug Resistance Situation of Streptococcus suis and Prevention Measures of Infections [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4920-4933.
[10]	WANG Jianing, ZHANG Ziqiang, KONG Dejing, FENG Caicai, ZHANG Feike, LIU Yumei. Isolation and Identification of Klebsiella pneumoniae in Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5198-5206.
[11]	LI Lili, CHEN Kaifeng, CHEN Bing, ZHOU Zhouping, WANG Nanwei, QU Xiaoyun, XU Chenggang, LIAO Ming, ZHANG Jianmin. Regulatory Role of STM1827 in the Biofilm Formation and Environmental Stress of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5207-5217.
[12]	ZHANG Kaichuan, WANG Jinyu, LI Shoujun, JIA Kun. Isolation, Identification and Biological Characteristics of Klebsiella pneumoniae from Sheep in Guangdong Province [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 328-337.
[13]	TONG Panpan, HUANG Shunmin, WANG Yudan, SHI Xuhui, CHEN Wenxia, SONG Xinlong, ZHANG Yi, SU Zhanqiang, XIE Jinxin. Phylogenetic Clustering, Serotype and Drug Resistance Analysis of Escherichia coli from Diarrhea with Piglets in Xinjiang [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 414-420.
[14]	CAO Li, CHENG Runan, WU Zhouhui, WANG Jiawei, WANG Yu, ZHANG Yonghong, WU Qingmin, WANG Zhen. Construction and Biological Characteristics of sapC Gene Deletion Strain of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2282-2289.
[15]	YU Yongfeng, QUAN Heng, DONG Wenhao, ZOU Ronghua, WU Xiaoni, GONG Xiaowei, CHEN Qiwei. The Mechanism of Two-component Regulatory System Mediating Drug Resistance of Gram-negative Bacteria [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1689-1701.