Deletion of the rstA gene of uropathogenic Escherichia coli strain U17 led to attenuation of virulence in mouse
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    Abstract:

    [Objective] To study the role of response regulator rstA of TCS RstA/RstB in pathogenesis of uropathogenic Escherichia coli (UPEC). [Methods] By using λ Red recombination system, we generated the rstA knockout mutant U17ΔrstA and the complementation strain ReU17ΔrstA. We then compared and analyzed the characterization of the mutant strain and the wild type strain in vivo and in vitro.[Results] The growth curves in LB showed that the deletion of rstA did not affect growth kinetics of mutant, whereas in LB-Fe medium, the growth rate of U17ΔrstA was lower than that of the wild-type strain U17. Under the selected environmental stress conditions in vitro, the bacterial survival experimental results showed that the mutant was not sensitive to acid, alkali, high osmotic pressure, urea and oxidants. Strain biofilm assay showed that the biofilm formation ability of the mutant was similar to that of the wild-type strain. qRT-PCR results showed that the rstA gene was significantly upregulated in LB-Fe medium, indicating that the iron-deficiency environment may be the stimulus signal evoking the RstA regulator. The mouse model of ascending urinary tract infection demonstrated that the deletion of rstA led to attenuation of virulence, because the mutant showed significantly decreased colonization compared with the wild type strain in urine, bladder and kidney, whereas the complementation strain restored the virulence to resemble that of wild-type strain. [Conclusion] The rstA gene was a potential virulence factor and associated with the pathogenesis of UPEC.

    Reference
    [1] Russo TA, Johnson JR. Proposal for a new inclusive designation for extraintestinal pathogenic isolates of Escherichia coli:ExPEC. The Journal of Infectious Diseases, 2000, 181(5):1753-1754.
    [2] Djorić D, Kristich CJ. Oxidative stress enhances cephalosporin resistance of Enterococcus faecalis through activation of a two-component signaling system. Antimicrobial Agents and Chemotherapy, 2015, 59(1):159-169.
    [3] Li Z, Fu Q, Wang Z, Li T, Zhang H, Guo F, Wang Y, Zhang J, Chen C. TceSR two-component regulatory system of Brucella melitensis 16M is involved in invasion, intracellular survival and regulated cytotoxicity for macrophages. Letters in Applied Microbiology, 2015, 60(6):565-571.
    [4] Zhang SM, Li XF, Wang X, Li Z, He J. The two-component signal transduction system YvcPQ regulates the bacterial resistance to bacitracin in Bacillus thuringiensis.Archives of Microbiology, 2016, 198(8):773-784.
    [5] Hoch JA. Two-component and phosphorelay signal transduction. Current Opinion in Microbiology, 2000, 3(2):165-170.
    [6] Yamamoto K, Hirao K, Oshima T, Aiba H, Utsumi R, Ishihama A. Functional characterization in vitro of all two-component signal transduction systems from Escherichia coli.Journal of Biological Chemistry, 2005, 280(2):1448-1456.
    [7] Zhou L, Lei XH, Bochner BR, Wanner BL. Phenotype microarray analysis of Escherichia coli K-12 mutants with deletions of all two-component systems. Journal of Bacteriology, 2003, 185(16):4956-4972.
    [8] Campbell TL, Ederer CS, Allali-Hassani A, Brown ED. Isolation of the rstA gene as a multicopy suppressor of YjeE, an essential ATPase of unknown function in Escherichia coli.Journal of Bacteriology, 2007, 189(8):3318-3321.
    [9] Ogasawara H, Hasegawa A, Kanda E, Miki T, Yamamoto K, Ishihama A. Genomic SELEX search for target promoters under the control of the PhoQP-RstBA signal relay cascade. Journal of Bacteriology, 2007, 189(13):4791-4799.
    [10] Cabeza ML, Aguirre A, Soncini FC, Véscovi EG. Induction of RpoS degradation by the two-component system regulator RstA in Salmonella enterica.Journal of Bacteriology, 2007, 189(20):7335-7342.
    [11] Tran TK, Han QQ, Shi YX, Guo L. A comparative proteomic analysis of Salmonella typhimurium under the regulation of the RstA/RstB and PhoP/PhoQ systems. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics, 2016, 1864(12):1686-1695.
    [12] Jeon J, Kim H, Yun J, Ryu S, Groisman EA, Shin D. RstA-promoted expression of the ferrous iron transporter FeoB under iron-replete conditions enhances Fur activity in Salmonella enterica.Journal of Bacteriology, 2008, 190(22):7326-7334.
    [13] Zhao LX, Gao S, Huan HX, Xu XJ, Zhu XP, Yang WX, Gao QQ, Liu XF. Comparison of virulence factors and expression of specific genes between uropathogenic Escherichia coli and avian pathogenic E.coli in a murine urinary tract infection model and a chicken challenge model. Microbiology, 2009, 155(5):1634-1644.
    [14] Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(12):6640-6645.
    [15] Večerek B, Moll I, Bläsi U. Control of Fur synthesis by the non-coding RNA RyhB and iron-responsive decoding. The EMBO Journal, 2007, 26(4):965-975.
    [16] Stepanović S, Vuković D, Dakić I, Savić B, Švabić-Vlahović M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods, 2000, 40(2):175-179.
    [17] Hagberg L, Engberg I, Freter R, Lam J, Olling S, Edén CS. Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin. Infection and Immunity, 1983, 40(1):273-283.
    [18] Haugen BJ, Pellett S, Redford P, Hamilton HL, Roesch PL, Welch RA. In vivo gene expression analysis identifies genes required for enhanced colonization of the mouse urinary tract by uropathogenic Escherichia coli strain CFT073dsdA.Infection and Immunity, 2007, 75(1):278-289.
    [19] Alteri CJ, Smith SN, Mobley HLT. Fitness of Escherichia coli during urinary tract infection requires gluconeogenesis and the TCA cycle. PLoS Pathogens, 2009, 5(5):e1000448.
    [20] Su Q, Guan TB, He Y, Lv HT. Siderophore biosynthesis governs the virulence of uropathogenic Escherichia coli by coordinately modulating the differential metabolism. Journal of Proteome Research, 2016, 15(4):1323-1332.
    [21] Kurabayashi K, Agata T, Asano H, Tomita H, Hirakawa H. Fur represses adhesion to, invasion of, and intracellular bacterial community formation within bladder epithelial cells and motility in uropathogenic Escherichia coli.Infection and Immunity, 2016, 84(22):3220-3231.
    [22] Tomenius H, Pernestig AK, Jonas K, Georgellis D, Möllby R, Normark S, Melefors Ö. The Escherichia coli BarA-UvrY two-component system is a virulence determinant in the urinary tract. BMC Microbiology, 2006, 6:27.
    [23] Hadjifrangiskou M, Kostakioti M, Chen SL, Henderson JP, Greene SE, Hultgren SJ. A central metabolic circuit controlled by QseC in pathogenic Escherichia coli.Molecular Microbiology, 2011, 80(6):1516-1529.
    [24] Alteri CJ, Lindner JR, Reiss DJ, Smith SN, Mobley HLT. The broadly conserved regulator PhoP links pathogen virulence and membrane potential in Escherichia coli.Molecular Microbiology, 2011, 82(1):145-163.
    [25] Pernestig AK, Normark SJ, Georgellis D, Melefors O. The role of the AirS two-component system in uropathogenic Escherichia coli.Advances in Experimental Medicine and Biology, 2000, 485:137-142.
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Qingqing Gao, Qiwen Shao, Zhengqin Ye, Le Xia, Song Gao, Xin'an Jiao, Xiufan Liu. Deletion of the rstA gene of uropathogenic Escherichia coli strain U17 led to attenuation of virulence in mouse. [J]. Acta Microbiologica Sinica, 2018, 58(3): 501-510

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History
  • Received:May 16,2017
  • Revised:May 31,2017
  • Online: February 12,2018
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