Cpx双组分调控系统在鳗弧菌应对环境压力和致病性中的作用
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摘要
Cpx双组分调控系统广泛存在于多种革兰氏阴性细菌,主要参与细菌膜压力变化的调控。作为信号感应器的组氨酸激酶CpxA感应外界环境信号,并传递给反应调节蛋白CpxR,后者作为转录调控因子调控靶基因表达,帮助细菌应对环境变化。
我们利用简并PCR和染色体步移技术,从鳗弧菌(Vibrio anguillarum)M3菌株的基因组克隆得到cpx基因簇,全长2768 bp。氨基酸序列分析表明,CpxR、CpxA与大肠杆菌和霍乱弧菌的相似性分别为60%和84%、45%和72%。
为了研究Cpx系统的功能,我们构建了cpxR和cpxA基因的非极性缺失突变株。表型特征分析显示,cpxR和cpxA的缺失对鳗弧菌的泳动、明胶酶活力、3.5%盐度下的生长能力没有明显影响(P >0.05),但是使鳗弧菌的菌膜形成能力减弱,对过氧化氢和SDS的敏感性增加,在pH9.0条件下的生长速度加快(P <0.05),对先锋霉素V和头孢呋肟的敏感性发生变化,表明鳗弧菌的Cpx系统参与了细菌对某些环境压力的感应和调控。我们以肌肉注射攻毒、浸泡攻毒和细菌体内存活分析等三个方面研究了突变株对蓝鳗龙致病能力的变化,结果显示,以注射途径进行攻毒,cpxR和cpxA缺失分别使细菌的半数致死量LD50提高了24.6倍和13.3倍,注射后突变株在鱼体内的存活能力都有所降低,而以浸泡途径进行攻毒,突变株和野生株的毒力没有明显变化,但是实验鱼的死亡时间延后,表明Cpx系统参与了鳗弧菌毒力的调控。
我们进一步用实时定量PCR(qRT-PCR)分析了cpxRA对10个毒力相关基因转录水平的影响,包括转录调控因子基因rpoS和rpoN,密度感应系统调控基因vanT,金属蛋白酶基因empA和prtV,胞外多糖合成和运输基因wza和wbfD,鞭毛蛋白基因flaA,六型分泌系统基因hcp和icmF。初步结果显示,上述基因的转录水平有一定变化,生物信息学分析初步表明,这些基因不是Cpx系统直接作用的靶基因,它们受到Cpx的影响尚待进一步研究证实。
Cpx two-component regulatory system exists in a variety of Gram-negative bacteria, mainly involved in the response to envelope stress. The sensor histidine kinase CpxA accepts signals from the environment, and passes them to the response regulator protein CpxR, which then functions as a transcription factor to regulate expression of target genes and helps bacteria respond to environmental changes. In this study,the cpx gene cluster was cloned from the genome DNA of Vibrio anguillarum M3 strain by degenerate PCR and genome walking. The obtained full length of cpx cluster was 2768 bp. The predicted CpxR and CpxA showed 60% and 84%, and 45% and 72% identities with Escherichia coli and Vibrio cholerae.
To investigate the function of cpxRA, the cpxR and cpxA in-frame deletion mutants were constructed from M3. The phenotypes of both mutants were comparatively analysed with the wild-type strain. Both cpxR and cpxA mutants showed no difference with the wild-type strain in the mobility, gelatinase activity, growth in TSB containing 3.5% NaCl (P >0.05), while both mutants showed reduced viability in 1% H2O2 and 0.05% SDS, reduced biofilm formation, increased growth ability at pH 9.0 (P <0.05), varied sensitivites to Cephazolin and Cefuroxime. These data indicated that V. anguillarum CpxRA participated in sensing environmental signals and regulating some specific stress responses.
Roles of cpxR and cpxA on bacterial virulence were determined. Bacteria were administrated to blue guorami fish Trichogaster trichopterus by intramuscular injection and immersing infection. In the injection route, the cpxR and cpxA mutant decreased 24.6-fold and 13.3-fold in virulence compared with the wild-type strain. In the immersion route, no differences in virulence were observed between both mutants and the wild-type, though, the death time of fish was delayed in the mutant strains. These data indicated that CpxRA was involved in the virulence of Vibrio anguillarum.
Then, transcription of ten virulence-related genes were analysed by real-time quantitative RT-PCR (qRT-PCR) in the mutants and the wild-type strains, including transcription regulator genes rpoS and rpoN, metalloproteinase genes empA and prtV, quorum sensing regulator vanT, exopolysaccharide biosythesis and transport genes wza, wbfD, flagellin A gene flaA, and type six secretion sytem genes hcp and icmF. A certain degree of changes in transcription of the ten genes were found between the mutants and the wild-type strains. Bioinformative analysis indicated that none of the ten genes were Cpx regulons. The mechanisms of V. anguillarum Cpx on these genes need further investigate.
我们利用简并PCR和染色体步移技术,从鳗弧菌(Vibrio anguillarum)M3菌株的基因组克隆得到cpx基因簇,全长2768 bp。氨基酸序列分析表明,CpxR、CpxA与大肠杆菌和霍乱弧菌的相似性分别为60%和84%、45%和72%。
为了研究Cpx系统的功能,我们构建了cpxR和cpxA基因的非极性缺失突变株。表型特征分析显示,cpxR和cpxA的缺失对鳗弧菌的泳动、明胶酶活力、3.5%盐度下的生长能力没有明显影响(P >0.05),但是使鳗弧菌的菌膜形成能力减弱,对过氧化氢和SDS的敏感性增加,在pH9.0条件下的生长速度加快(P <0.05),对先锋霉素V和头孢呋肟的敏感性发生变化,表明鳗弧菌的Cpx系统参与了细菌对某些环境压力的感应和调控。我们以肌肉注射攻毒、浸泡攻毒和细菌体内存活分析等三个方面研究了突变株对蓝鳗龙致病能力的变化,结果显示,以注射途径进行攻毒,cpxR和cpxA缺失分别使细菌的半数致死量LD50提高了24.6倍和13.3倍,注射后突变株在鱼体内的存活能力都有所降低,而以浸泡途径进行攻毒,突变株和野生株的毒力没有明显变化,但是实验鱼的死亡时间延后,表明Cpx系统参与了鳗弧菌毒力的调控。
我们进一步用实时定量PCR(qRT-PCR)分析了cpxRA对10个毒力相关基因转录水平的影响,包括转录调控因子基因rpoS和rpoN,密度感应系统调控基因vanT,金属蛋白酶基因empA和prtV,胞外多糖合成和运输基因wza和wbfD,鞭毛蛋白基因flaA,六型分泌系统基因hcp和icmF。初步结果显示,上述基因的转录水平有一定变化,生物信息学分析初步表明,这些基因不是Cpx系统直接作用的靶基因,它们受到Cpx的影响尚待进一步研究证实。
Cpx two-component regulatory system exists in a variety of Gram-negative bacteria, mainly involved in the response to envelope stress. The sensor histidine kinase CpxA accepts signals from the environment, and passes them to the response regulator protein CpxR, which then functions as a transcription factor to regulate expression of target genes and helps bacteria respond to environmental changes. In this study,the cpx gene cluster was cloned from the genome DNA of Vibrio anguillarum M3 strain by degenerate PCR and genome walking. The obtained full length of cpx cluster was 2768 bp. The predicted CpxR and CpxA showed 60% and 84%, and 45% and 72% identities with Escherichia coli and Vibrio cholerae.
To investigate the function of cpxRA, the cpxR and cpxA in-frame deletion mutants were constructed from M3. The phenotypes of both mutants were comparatively analysed with the wild-type strain. Both cpxR and cpxA mutants showed no difference with the wild-type strain in the mobility, gelatinase activity, growth in TSB containing 3.5% NaCl (P >0.05), while both mutants showed reduced viability in 1% H2O2 and 0.05% SDS, reduced biofilm formation, increased growth ability at pH 9.0 (P <0.05), varied sensitivites to Cephazolin and Cefuroxime. These data indicated that V. anguillarum CpxRA participated in sensing environmental signals and regulating some specific stress responses.
Roles of cpxR and cpxA on bacterial virulence were determined. Bacteria were administrated to blue guorami fish Trichogaster trichopterus by intramuscular injection and immersing infection. In the injection route, the cpxR and cpxA mutant decreased 24.6-fold and 13.3-fold in virulence compared with the wild-type strain. In the immersion route, no differences in virulence were observed between both mutants and the wild-type, though, the death time of fish was delayed in the mutant strains. These data indicated that CpxRA was involved in the virulence of Vibrio anguillarum.
Then, transcription of ten virulence-related genes were analysed by real-time quantitative RT-PCR (qRT-PCR) in the mutants and the wild-type strains, including transcription regulator genes rpoS and rpoN, metalloproteinase genes empA and prtV, quorum sensing regulator vanT, exopolysaccharide biosythesis and transport genes wza, wbfD, flagellin A gene flaA, and type six secretion sytem genes hcp and icmF. A certain degree of changes in transcription of the ten genes were found between the mutants and the wild-type strains. Bioinformative analysis indicated that none of the ten genes were Cpx regulons. The mechanisms of V. anguillarum Cpx on these genes need further investigate.
引文
Alm E, Huang K, and Arkin A. The evolution of two-component systems in bacteria reveals different strategies for niche adaptation[J]. Plos Comput Biol. 2006, 2 (11): 1329-1342
Altman E,Segal G. The response regulator CpxR directly regulates expression of several Legionella pneumophila icm/dot components as well as new translocated substrates[J]. J Bacteriol. 2008, 190 (6): 1985-1996
Austin B,Allen-Austin D. Bacterial pathogens of fish[J]. J Appl Bacteriol. 1985, 58 (5): 483-506
Carlsson K E, Liu J, Edqvist P J, et al. Influence of the Cpx extracytoplasmic-stress-responsive pathway on Yersinia sp.-eukaryotic cell contact[J]. Infect Immun. 2007, 75 (9): 4386-4399
Chen Q, Wertheimer A M, Tolmasky M E, et al. The AngR protein and the siderophore anguibactin positively regulate the expression of iron-transport genes in Vibrio anguillarum[J]. Mol Microbiol. 1996, 22 (1): 127-134
Conrad M. Cross-scale information processing in evolution, development and intelligence[J]. Biosystems. 1996, 38 (2-3): 97-109
Cozzone A J. Protein phosphorylation in prokaryotes[J]. Annu Rev Microbiol. 1988, 42: 97-125
Croxatto A, Chalker V J, Lauritz J, et al. VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum[J]. J Bacteriol. 2002, 184 (6): 1617-1629
Croxatto A, Lauritz J, Chen C, et al. Vibrio anguillarum colonization of rainbow trout integument requires a DNA locus involved in exopolysaccharide transport and biosynthesis[J]. Environ Microbiol. 2007, 9 (2): 370-382
Croxatto A, Pride J, Hardman A, et al. A distinctive dual-channel quorum-sensing system operates in Vibrio anguillarum[J]. Mol Microbiol. 2004, 52 (6): 1677-1689
Danese P N,Silhavy T J. The sigma(E) and the Cpx signal transduction systems control the synthesis of periplasmic protein-folding enzymes in Escherichia coli[J]. Genes Dev. 1997, 11 (9): 1183-1193
Danese P N,Silhavy T J. CpxP, a stress-combative member of the Cpx regulon[J]. J Bacteriol. 1998, 180 (4): 831-839
Danese P N, Snyder W B, Cosma C L, et al. The Cpx two-component signal transduction pathway of Escherichia coli regulates transcription of the gene specifying the stress-inducible periplasmic protease, DegP[J]. Genes Dev. 1995, 9 (4): 387-398
Dartigalongue C,Raina S. A new heat-shock gene, ppiD, encodes a peptidyl-prolyl isomerase required for folding of outer membrane proteins in Escherichia coli[J]. EMBO J. 1998, 17 (14): 3968-3980
Dong J M, Iuchi S, Kwan H S, et al. The deduced amino-acid-sequence of the cloned cpxR gene suggests the protein is the cognate regulator for the membrane sensor, CpxA, in a 2-component signal-transduction system of Escherichia coli[J]. Gene. 1993, 136 (1-2): 227-230
Dorel C, Vidal O, Prigent-Combaret C, et al. Involvement of the Cpx signal transduction pathway of E. coli in biofilm formation[J]. FEMS Microbiol Lett. 1999, 178 (1): 169-175
Edwards R A, Keller L H, and Schifferli D M. Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression[J]. Gene. 1998, 207 (2): 149-157
Filloux A, Hachani A, and Bleves S. The bacterial type VI secretion machine: yet another player for protein transport across membranes[J]. Microbiology-Sgm. 2008, 154: 1570-1583
Gal-Mor O,Segal G. Identification of CpxR as a positive regulator of icm and dot virulence genes of Legionella pneumophila[J]. J Bacteriol. 2003, 185 (16): 4908-4919
Galperin M Y. A census of membrane-bound and intracellular signal transduction proteins in bacteria: bacterial IQ, extroverts and introverts[J]. Bmc Microbiol. 2005, 5: 35
Goodman A L, Merighi M, Hyodo M, et al. Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen[J]. Genes Dev. 2009, 23 (2): 249-259
Grigoroudis A I, Panagiotidis C A, Lioliou E E, et al. Molecular modeling and functional analysis of the AtoS-AtoC two-component signal transduction system of Escherichia coli[J]. Biochim Biophys Acta. 2007, 1770 (8): 1248-1258
Hagiwara D, Yamashino T, and Mizuno T. A genome-wide view of the Escherichia coli BasS-BasR two-component system implicated in iron-responses[J]. Biosci Biotech Bioch. 2004, 68 (8): 1758-1767
Herbert E E, Cowles K N, and Goodrich-Blair H. CpxRA regulates mutualism and pathogenesis in Xenorhabdus nematophila[J]. Appl Environ Microbiol. 2007, 73 (24): 7826-7836
Hess J F, Oosawa K, Matsumura P, et al. Protein phosphorylation is involved in bacterial chemotaxis[J]. Proc Natl Acad Sci U S A. 1987, 84 (21): 7609-7613
Hirakawa H, Nishino K, Hirata T, et al. Comprehensive studies of drug resistance mediated by overexpression of response regulators of two-component signal transduction systems in Escherichia coli[J]. J Bacteriol. 2003, 185 (6): 1851-1856
Ho S N, Hunt H D, Horton R M, et al. Site-directed mutagenesis by overlap extension using the polymerase chain-reaction[J]. Gene. 1989, 77 (1): 51-59
Hu Y H, Liu C S, Hou J H, et al. Identification, characterization, and molecular application of a virulence-associated autotransporter from a pathogenic Pseudomonas fluorescens strain[J]. Appl Environ Microbiol. 2009, 75 (13): 4333-4340
Humphreys S, Rowley G, Stevenson A, et al. Role of the two-component regulator CpxAR in the virulence of Salmonella entetica serotype typhimurium[J]. Infection and Immunity. 2004, 72 (8): 4654-4661
Hung D L, Raivio T L, Jones C H, et al. Cpx signaling pathway monitors biogenesis and affects assembly and expression of P pili[J]. EMBO J. 2001, 20 (7): 1508-1518
Isaac D D, Pinkner J S, Hultgren S J, et al. The extracytoplasmic adaptor protein CpxP is degraded with substrate by DegP[J]. Proc Natl Acad Sci U S A. 2005, 102 (49): 17775-17779
Jubelin G, Vianney A, Beloin C, et al. CpxR/OmpR interplay regulates curli gene expression in response to osmolarity in Escherichia coli[J]. J Bacteriol. 2005, 187 (6): 2038-2049
Kim S W, Moon K H, Baik H S, et al. Changes of physiological and biochemical properties of Salmonella enterica serovar Typhimurium by deletion of cpxR and lon genes using allelic exchange method[J]. J Microbiol Methods. 2009, 79 (3): 314-320
Kirby J R. Chemotaxis-like regulatory systems: unique roles in diverse bacteria[J]. Annu Rev Microbiol. 2009, 63: 45-59
Lan M Z, Peng X, Xiang M Y, et al. Construction and characterization of a live, attenuated esrB mutant of Edwardsiella tarda and its potential as a vaccine against the haemorrhagic septicaemia in turbot, Scophthamus maximus (L.)[J]. Fish Shellfish Immunol. 2007, 23 (3): 521-530
Li L, Rock J L, and Nelson D R. Identification and characterization of a repeat-in-toxin gene cluster in Vibrio anguillarum[J]. Infect Immun. 2008, 76 (6): 2620-2632
Lucana D O D,Groves M R. The three-component signalling system HbpS-SenS-SenR as an example of a redox sensing pathway in bacteria[J]. Amino Acids. 2009, 37 (3): 479-486
Marijuán P C, Navarro J, and del Moral R. On prokaryotic intelligence: Strategies for sensing the environment[J]. Biosystems. 2010, 99 (2): 94-103
Martin C, Williams A, Hernandez-Pando R, et al. The live Mycobacterium tuberculosis phoP mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs[J]. Vaccine. 2006, 24 (17): 3408-3419
Matsuda K, Chaudhari A A, Kim S W, et al. Physiology, pathogenicity and immunogenicity of lon and/or cpxR deleted mutants of Salmonella Gallinarum as vaccine candidates for fowl typhoid[J]. Vet Res. 2010, 41 (5): 59
McGee K, Horstedt P, and Milton D L. Identification and characterization of additional flagellin genes from Vibrio anguillarum[J]. J Bacteriol. 1996, 178 (17): 5188-5198
Milton D L, Norqvist A, and Wolf-Watz H. Cloning of a metalloprotease gene involved in the virulence mechanism of Vibrio anguillarum[J]. J Bacteriol. 1992, 174 (22): 7235-7244
Milton D L, Norqvist A, and Wolfwatz H. Sequence of a Novel Virulence-Mediating Gene, Virc, from Vibrio-Anguillarum[J]. Gene. 1995, 164 (1): 95-100
Milton D L, O'Toole R, Horstedt P, et al. Flagellin A is essential for the virulence of Vibrio anguillarum[J]. J Bacteriol. 1996, 178 (5): 1310-1319
Mo Z L, Guo D S, Mao Y X, et al. Identification and characterization of the Vibrio anguillarum prtV gene encoding a new metalloprotease[J]. Chin J Oceanol Limn. 2010, 28 (1): 55-61
Nakayama S,Watanabe H. Involvement of cpxA, a sensor of a two-component regulatory system, in the pH-dependent regulation of expression of Shigella sonnei virF gene[J]. J Bacteriol. 1995, 177 (17): 5062-5069
Nevesinjac A Z,Raivio T L. The Cpx envelope stress response affects expression of the type IV bundle-forming pili of enteropathogenic Escherichia coli[J]. J Bacteriol. 2005, 187 (2): 672-686
Ninfa A J,Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli[J]. Proc Natl Acad Sci U S A. 1986, 83 (16): 5909-5913
Nixon B T, Ronson C W, and Ausubel F M. Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC[J]. Proc Natl Acad Sci U S A. 1986, 83 (20): 7850-7854
Norqvist A, Norrman B, and Wolf-Watz H. Identification and characterization of a zinc metalloprotease associated with invasion by the fish pathogen Vibrio anguillarum[J]. Infect Immun. 1990, 58 (11): 3731-3736
O'Toole R, Milton D L, Horstedt P, et al. RpoN of the fish pathogen Vibrio (Listonella) anguillarum is essential for flagellum production and virulence by the water-borne but not intraperitoneal route of inoculation[J]. Microbiology-Sgm. 1997, 143: 3849-3859
Ormonde P, Horstedt P, O'Toole R, et al. Role of motility in adherence to and invasion of a fish cell line by Vibrio anguillarum[J]. J Bacteriol. 2000, 182 (8): 2326-2328
OToole R, Milton D L, and WolfWatz H. Chemotactic motility is required for invasion of the host by the fish pathogen Vibrio anguillarum[J]. Molecular Microbiology. 1996, 19 (3): 625-637
Pogliano J, Lynch A S, Belin D, et al. Regulation of Escherichia coli cell envelope proteins involved in protein folding and degradation by the Cpx two-component system[J]. Genes Dev. 1997, 11 (9): 1169-1182
Prigent-Combaret C, Brombacher E, Vidal O, et al. Complex regulatory network controls initial adhesion and biofilm formation in Escherichia coli via regulation of the csgD gene[J]. J Bacteriol. 2001, 183 (24): 7213-7223
Raffa R G,Raivio T L. A third envelope stress signal transduction pathway in Escherichia coli[J]. Mol Microbiol. 2002, 45 (6): 1599-1611
Raivio T L, Popkin D L, and Silhavy T J. The Cpx envelope stress response is controlled by amplification and feedback inhibition[J]. J Bacteriol. 1999a, 181 (17): 5263-5272
Raivio T L,Silhavy T J. Transduction of envelope stress in Escherichia coli by the Cpx two-component system[J]. J Bacteriol. 1997, 179 (24): 7724-7733
Raivio T L,Silhavy T J. The sigmaE and Cpx regulatory pathways: overlapping but distinct envelope stress responses[J]. Curr Opin Microbiol. 1999b, 2 (2): 159-165
Rodkhum C, Hirono I, Crosa J H, et al. Four novel hemolysin genes of Vibrio anguillarum and their virulence to rainbow trout[J]. Microb Pathog. 2005, 39 (4): 109-119
Rodkhum C, Hirono I, Stork M, et al. Putative virulence-related genes in Vibrio anguillarum identified by random genome sequencing[J]. J Fish Dis. 2006, 29 (3): 157-166
Rodrigue A, Quentin Y, Lazdunski A, et al. Two-component systems in Pseudomonas aeruginosa: why so many?[J]. Trends Microbiol. 2000, 8 (11): 498-504
Rubires X, Saigi F, Pique N, et al. A gene (wbbL) from Serratia marcescens N28b (O4) complements the rfb-50 mutation of Escherichia coli K-12 derivatives[J]. J Bacteriol. 1997, 179 (23): 7581-7586
Salinas P C,Crosa J H. Regulation of angR, a gene with regulatory and biosynthetic functions in the pJM1 plasmid-mediated iron uptake system of Vibrio anguillarum[J]. Gene. 1995, 160 (1): 17-23
Scharf B E, Aldridge P D, Kirby J R, et al. Upward mobility and alternative lifestyles: a report from the 10th biennial meeting on Bacterial Locomotion and Signal Transduction[J]. Mol Microbiol. 2009, 73 (1): 5-19
Slamti L,Waldor M K. Genetic analysis of activation of the Vibrio cholerae Cpx pathway[J]. J Bacteriol. 2009, 191 (16): 5044-5056
Spinola S M, Fortney K R, Baker B, et al. Activation of the CpxRA System by Deletion of cpxA Impairs the Ability of Haemophilus ducreyi to Infect Humans[J]. Infect Immun. 2010, 78 (9): 3898-3904
Staroscik A M, Denkin S M, and Nelson D R. Regulation of the Vibrio anguillarum metalloprotease EmpA by posttranslational modification[J]. J Bacteriol. 2005, 187 (7): 2257-2260
Stork M, Di Lorenzo M, Mourino S, et al. Two tonB systems function in iron transport in Vibrio anguillarum, but only one is essential for virulence[J]. Infect Immun. 2004, 72 (12): 7326-7329
Stork M, Di Lorenzo M, Welch T J, et al. Plasmid-mediated iron uptake and virulence in Vibrio anguillarum[J]. Plasmid. 2002, 48 (3): 222-228
Thompson F L, Iida T, and Swings J. Biodiversity of vibrios[J]. Microbiol Mol Biol Rev. 2004, 68 (3): 403-431
Ulrich L E, Koonin E V, and Zhulin I B. One-component systems dominate signal transduction in prokaryotes[J]. Trends Microbiol. 2005, 13 (2): 52-56
Wang S Y, Lauritz J, Jass J, et al. Role for the major outer-membrane protein from Vibrio anguillarum in bile resistance and biofilm formation[J]. Microbiology. 2003, 149 (Pt 4): 1061-1071
Weatherspoon-Griffin N, Zhao G, Kong W, et al. The CpxR/CpxA two-component system up-regulates two Tat-dependent peptidoglycan amidases to confer bacterial resistance to antimicrobial peptide[J]. J Biol Chem. 2011, 286 (7): 5529-5539
Weber B, Croxatto A, Chen C, et al. RpoS induces expression of the Vibrio anguillarum quorum-sensing regulator VanT[J]. Microbiology. 2008, 154 (Pt 3): 767-780
Weber B, Hasic M, Chen C, et al. Type VI secretion modulates quorum sensing and stress response in Vibrio anguillarum[J]. Environ Microbiol. 2009, 11 (12): 3018-3028
Weber R F,Silverman P M. The Cpx proteins of Escherichia coli K12: structure of the CpxA polypeptide as an inner membrane component[J]. Journal of Molecular Biology. 1988, 203 (2): 467-478
Whitfield C. Biosynthesis and assembly of capsular polysaccharides in Escherichia coli[J]. Annu Rev Biochem. 2006, 75: 39-68
Xiao P, Mo Z L, Mao Y X, et al. Detection of Vibrio anguillarum by PCR amplification of the empA gene[J]. Journal of Fish Diseases. 2009, 32 (3): 293-296
莫照兰,茅云翔,陈师勇,等.一株牙鲆皮肤溃烂症病原菌的鉴定[J].微生物学报. 2002, (03): 263-269
杨少丽,王印庚,董树刚.海水养殖鱼类弧菌病的研究进展[J].海洋水产研究. 2005, (04): 75-83
杨茂成.兽医统计学[M].北京:中国展望出版社, 1990: 232-234.
Altman E,Segal G. The response regulator CpxR directly regulates expression of several Legionella pneumophila icm/dot components as well as new translocated substrates[J]. J Bacteriol. 2008, 190 (6): 1985-1996
Austin B,Allen-Austin D. Bacterial pathogens of fish[J]. J Appl Bacteriol. 1985, 58 (5): 483-506
Carlsson K E, Liu J, Edqvist P J, et al. Influence of the Cpx extracytoplasmic-stress-responsive pathway on Yersinia sp.-eukaryotic cell contact[J]. Infect Immun. 2007, 75 (9): 4386-4399
Chen Q, Wertheimer A M, Tolmasky M E, et al. The AngR protein and the siderophore anguibactin positively regulate the expression of iron-transport genes in Vibrio anguillarum[J]. Mol Microbiol. 1996, 22 (1): 127-134
Conrad M. Cross-scale information processing in evolution, development and intelligence[J]. Biosystems. 1996, 38 (2-3): 97-109
Cozzone A J. Protein phosphorylation in prokaryotes[J]. Annu Rev Microbiol. 1988, 42: 97-125
Croxatto A, Chalker V J, Lauritz J, et al. VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum[J]. J Bacteriol. 2002, 184 (6): 1617-1629
Croxatto A, Lauritz J, Chen C, et al. Vibrio anguillarum colonization of rainbow trout integument requires a DNA locus involved in exopolysaccharide transport and biosynthesis[J]. Environ Microbiol. 2007, 9 (2): 370-382
Croxatto A, Pride J, Hardman A, et al. A distinctive dual-channel quorum-sensing system operates in Vibrio anguillarum[J]. Mol Microbiol. 2004, 52 (6): 1677-1689
Danese P N,Silhavy T J. The sigma(E) and the Cpx signal transduction systems control the synthesis of periplasmic protein-folding enzymes in Escherichia coli[J]. Genes Dev. 1997, 11 (9): 1183-1193
Danese P N,Silhavy T J. CpxP, a stress-combative member of the Cpx regulon[J]. J Bacteriol. 1998, 180 (4): 831-839
Danese P N, Snyder W B, Cosma C L, et al. The Cpx two-component signal transduction pathway of Escherichia coli regulates transcription of the gene specifying the stress-inducible periplasmic protease, DegP[J]. Genes Dev. 1995, 9 (4): 387-398
Dartigalongue C,Raina S. A new heat-shock gene, ppiD, encodes a peptidyl-prolyl isomerase required for folding of outer membrane proteins in Escherichia coli[J]. EMBO J. 1998, 17 (14): 3968-3980
Dong J M, Iuchi S, Kwan H S, et al. The deduced amino-acid-sequence of the cloned cpxR gene suggests the protein is the cognate regulator for the membrane sensor, CpxA, in a 2-component signal-transduction system of Escherichia coli[J]. Gene. 1993, 136 (1-2): 227-230
Dorel C, Vidal O, Prigent-Combaret C, et al. Involvement of the Cpx signal transduction pathway of E. coli in biofilm formation[J]. FEMS Microbiol Lett. 1999, 178 (1): 169-175
Edwards R A, Keller L H, and Schifferli D M. Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression[J]. Gene. 1998, 207 (2): 149-157
Filloux A, Hachani A, and Bleves S. The bacterial type VI secretion machine: yet another player for protein transport across membranes[J]. Microbiology-Sgm. 2008, 154: 1570-1583
Gal-Mor O,Segal G. Identification of CpxR as a positive regulator of icm and dot virulence genes of Legionella pneumophila[J]. J Bacteriol. 2003, 185 (16): 4908-4919
Galperin M Y. A census of membrane-bound and intracellular signal transduction proteins in bacteria: bacterial IQ, extroverts and introverts[J]. Bmc Microbiol. 2005, 5: 35
Goodman A L, Merighi M, Hyodo M, et al. Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen[J]. Genes Dev. 2009, 23 (2): 249-259
Grigoroudis A I, Panagiotidis C A, Lioliou E E, et al. Molecular modeling and functional analysis of the AtoS-AtoC two-component signal transduction system of Escherichia coli[J]. Biochim Biophys Acta. 2007, 1770 (8): 1248-1258
Hagiwara D, Yamashino T, and Mizuno T. A genome-wide view of the Escherichia coli BasS-BasR two-component system implicated in iron-responses[J]. Biosci Biotech Bioch. 2004, 68 (8): 1758-1767
Herbert E E, Cowles K N, and Goodrich-Blair H. CpxRA regulates mutualism and pathogenesis in Xenorhabdus nematophila[J]. Appl Environ Microbiol. 2007, 73 (24): 7826-7836
Hess J F, Oosawa K, Matsumura P, et al. Protein phosphorylation is involved in bacterial chemotaxis[J]. Proc Natl Acad Sci U S A. 1987, 84 (21): 7609-7613
Hirakawa H, Nishino K, Hirata T, et al. Comprehensive studies of drug resistance mediated by overexpression of response regulators of two-component signal transduction systems in Escherichia coli[J]. J Bacteriol. 2003, 185 (6): 1851-1856
Ho S N, Hunt H D, Horton R M, et al. Site-directed mutagenesis by overlap extension using the polymerase chain-reaction[J]. Gene. 1989, 77 (1): 51-59
Hu Y H, Liu C S, Hou J H, et al. Identification, characterization, and molecular application of a virulence-associated autotransporter from a pathogenic Pseudomonas fluorescens strain[J]. Appl Environ Microbiol. 2009, 75 (13): 4333-4340
Humphreys S, Rowley G, Stevenson A, et al. Role of the two-component regulator CpxAR in the virulence of Salmonella entetica serotype typhimurium[J]. Infection and Immunity. 2004, 72 (8): 4654-4661
Hung D L, Raivio T L, Jones C H, et al. Cpx signaling pathway monitors biogenesis and affects assembly and expression of P pili[J]. EMBO J. 2001, 20 (7): 1508-1518
Isaac D D, Pinkner J S, Hultgren S J, et al. The extracytoplasmic adaptor protein CpxP is degraded with substrate by DegP[J]. Proc Natl Acad Sci U S A. 2005, 102 (49): 17775-17779
Jubelin G, Vianney A, Beloin C, et al. CpxR/OmpR interplay regulates curli gene expression in response to osmolarity in Escherichia coli[J]. J Bacteriol. 2005, 187 (6): 2038-2049
Kim S W, Moon K H, Baik H S, et al. Changes of physiological and biochemical properties of Salmonella enterica serovar Typhimurium by deletion of cpxR and lon genes using allelic exchange method[J]. J Microbiol Methods. 2009, 79 (3): 314-320
Kirby J R. Chemotaxis-like regulatory systems: unique roles in diverse bacteria[J]. Annu Rev Microbiol. 2009, 63: 45-59
Lan M Z, Peng X, Xiang M Y, et al. Construction and characterization of a live, attenuated esrB mutant of Edwardsiella tarda and its potential as a vaccine against the haemorrhagic septicaemia in turbot, Scophthamus maximus (L.)[J]. Fish Shellfish Immunol. 2007, 23 (3): 521-530
Li L, Rock J L, and Nelson D R. Identification and characterization of a repeat-in-toxin gene cluster in Vibrio anguillarum[J]. Infect Immun. 2008, 76 (6): 2620-2632
Lucana D O D,Groves M R. The three-component signalling system HbpS-SenS-SenR as an example of a redox sensing pathway in bacteria[J]. Amino Acids. 2009, 37 (3): 479-486
Marijuán P C, Navarro J, and del Moral R. On prokaryotic intelligence: Strategies for sensing the environment[J]. Biosystems. 2010, 99 (2): 94-103
Martin C, Williams A, Hernandez-Pando R, et al. The live Mycobacterium tuberculosis phoP mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs[J]. Vaccine. 2006, 24 (17): 3408-3419
Matsuda K, Chaudhari A A, Kim S W, et al. Physiology, pathogenicity and immunogenicity of lon and/or cpxR deleted mutants of Salmonella Gallinarum as vaccine candidates for fowl typhoid[J]. Vet Res. 2010, 41 (5): 59
McGee K, Horstedt P, and Milton D L. Identification and characterization of additional flagellin genes from Vibrio anguillarum[J]. J Bacteriol. 1996, 178 (17): 5188-5198
Milton D L, Norqvist A, and Wolf-Watz H. Cloning of a metalloprotease gene involved in the virulence mechanism of Vibrio anguillarum[J]. J Bacteriol. 1992, 174 (22): 7235-7244
Milton D L, Norqvist A, and Wolfwatz H. Sequence of a Novel Virulence-Mediating Gene, Virc, from Vibrio-Anguillarum[J]. Gene. 1995, 164 (1): 95-100
Milton D L, O'Toole R, Horstedt P, et al. Flagellin A is essential for the virulence of Vibrio anguillarum[J]. J Bacteriol. 1996, 178 (5): 1310-1319
Mo Z L, Guo D S, Mao Y X, et al. Identification and characterization of the Vibrio anguillarum prtV gene encoding a new metalloprotease[J]. Chin J Oceanol Limn. 2010, 28 (1): 55-61
Nakayama S,Watanabe H. Involvement of cpxA, a sensor of a two-component regulatory system, in the pH-dependent regulation of expression of Shigella sonnei virF gene[J]. J Bacteriol. 1995, 177 (17): 5062-5069
Nevesinjac A Z,Raivio T L. The Cpx envelope stress response affects expression of the type IV bundle-forming pili of enteropathogenic Escherichia coli[J]. J Bacteriol. 2005, 187 (2): 672-686
Ninfa A J,Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli[J]. Proc Natl Acad Sci U S A. 1986, 83 (16): 5909-5913
Nixon B T, Ronson C W, and Ausubel F M. Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC[J]. Proc Natl Acad Sci U S A. 1986, 83 (20): 7850-7854
Norqvist A, Norrman B, and Wolf-Watz H. Identification and characterization of a zinc metalloprotease associated with invasion by the fish pathogen Vibrio anguillarum[J]. Infect Immun. 1990, 58 (11): 3731-3736
O'Toole R, Milton D L, Horstedt P, et al. RpoN of the fish pathogen Vibrio (Listonella) anguillarum is essential for flagellum production and virulence by the water-borne but not intraperitoneal route of inoculation[J]. Microbiology-Sgm. 1997, 143: 3849-3859
Ormonde P, Horstedt P, O'Toole R, et al. Role of motility in adherence to and invasion of a fish cell line by Vibrio anguillarum[J]. J Bacteriol. 2000, 182 (8): 2326-2328
OToole R, Milton D L, and WolfWatz H. Chemotactic motility is required for invasion of the host by the fish pathogen Vibrio anguillarum[J]. Molecular Microbiology. 1996, 19 (3): 625-637
Pogliano J, Lynch A S, Belin D, et al. Regulation of Escherichia coli cell envelope proteins involved in protein folding and degradation by the Cpx two-component system[J]. Genes Dev. 1997, 11 (9): 1169-1182
Prigent-Combaret C, Brombacher E, Vidal O, et al. Complex regulatory network controls initial adhesion and biofilm formation in Escherichia coli via regulation of the csgD gene[J]. J Bacteriol. 2001, 183 (24): 7213-7223
Raffa R G,Raivio T L. A third envelope stress signal transduction pathway in Escherichia coli[J]. Mol Microbiol. 2002, 45 (6): 1599-1611
Raivio T L, Popkin D L, and Silhavy T J. The Cpx envelope stress response is controlled by amplification and feedback inhibition[J]. J Bacteriol. 1999a, 181 (17): 5263-5272
Raivio T L,Silhavy T J. Transduction of envelope stress in Escherichia coli by the Cpx two-component system[J]. J Bacteriol. 1997, 179 (24): 7724-7733
Raivio T L,Silhavy T J. The sigmaE and Cpx regulatory pathways: overlapping but distinct envelope stress responses[J]. Curr Opin Microbiol. 1999b, 2 (2): 159-165
Rodkhum C, Hirono I, Crosa J H, et al. Four novel hemolysin genes of Vibrio anguillarum and their virulence to rainbow trout[J]. Microb Pathog. 2005, 39 (4): 109-119
Rodkhum C, Hirono I, Stork M, et al. Putative virulence-related genes in Vibrio anguillarum identified by random genome sequencing[J]. J Fish Dis. 2006, 29 (3): 157-166
Rodrigue A, Quentin Y, Lazdunski A, et al. Two-component systems in Pseudomonas aeruginosa: why so many?[J]. Trends Microbiol. 2000, 8 (11): 498-504
Rubires X, Saigi F, Pique N, et al. A gene (wbbL) from Serratia marcescens N28b (O4) complements the rfb-50 mutation of Escherichia coli K-12 derivatives[J]. J Bacteriol. 1997, 179 (23): 7581-7586
Salinas P C,Crosa J H. Regulation of angR, a gene with regulatory and biosynthetic functions in the pJM1 plasmid-mediated iron uptake system of Vibrio anguillarum[J]. Gene. 1995, 160 (1): 17-23
Scharf B E, Aldridge P D, Kirby J R, et al. Upward mobility and alternative lifestyles: a report from the 10th biennial meeting on Bacterial Locomotion and Signal Transduction[J]. Mol Microbiol. 2009, 73 (1): 5-19
Slamti L,Waldor M K. Genetic analysis of activation of the Vibrio cholerae Cpx pathway[J]. J Bacteriol. 2009, 191 (16): 5044-5056
Spinola S M, Fortney K R, Baker B, et al. Activation of the CpxRA System by Deletion of cpxA Impairs the Ability of Haemophilus ducreyi to Infect Humans[J]. Infect Immun. 2010, 78 (9): 3898-3904
Staroscik A M, Denkin S M, and Nelson D R. Regulation of the Vibrio anguillarum metalloprotease EmpA by posttranslational modification[J]. J Bacteriol. 2005, 187 (7): 2257-2260
Stork M, Di Lorenzo M, Mourino S, et al. Two tonB systems function in iron transport in Vibrio anguillarum, but only one is essential for virulence[J]. Infect Immun. 2004, 72 (12): 7326-7329
Stork M, Di Lorenzo M, Welch T J, et al. Plasmid-mediated iron uptake and virulence in Vibrio anguillarum[J]. Plasmid. 2002, 48 (3): 222-228
Thompson F L, Iida T, and Swings J. Biodiversity of vibrios[J]. Microbiol Mol Biol Rev. 2004, 68 (3): 403-431
Ulrich L E, Koonin E V, and Zhulin I B. One-component systems dominate signal transduction in prokaryotes[J]. Trends Microbiol. 2005, 13 (2): 52-56
Wang S Y, Lauritz J, Jass J, et al. Role for the major outer-membrane protein from Vibrio anguillarum in bile resistance and biofilm formation[J]. Microbiology. 2003, 149 (Pt 4): 1061-1071
Weatherspoon-Griffin N, Zhao G, Kong W, et al. The CpxR/CpxA two-component system up-regulates two Tat-dependent peptidoglycan amidases to confer bacterial resistance to antimicrobial peptide[J]. J Biol Chem. 2011, 286 (7): 5529-5539
Weber B, Croxatto A, Chen C, et al. RpoS induces expression of the Vibrio anguillarum quorum-sensing regulator VanT[J]. Microbiology. 2008, 154 (Pt 3): 767-780
Weber B, Hasic M, Chen C, et al. Type VI secretion modulates quorum sensing and stress response in Vibrio anguillarum[J]. Environ Microbiol. 2009, 11 (12): 3018-3028
Weber R F,Silverman P M. The Cpx proteins of Escherichia coli K12: structure of the CpxA polypeptide as an inner membrane component[J]. Journal of Molecular Biology. 1988, 203 (2): 467-478
Whitfield C. Biosynthesis and assembly of capsular polysaccharides in Escherichia coli[J]. Annu Rev Biochem. 2006, 75: 39-68
Xiao P, Mo Z L, Mao Y X, et al. Detection of Vibrio anguillarum by PCR amplification of the empA gene[J]. Journal of Fish Diseases. 2009, 32 (3): 293-296
莫照兰,茅云翔,陈师勇,等.一株牙鲆皮肤溃烂症病原菌的鉴定[J].微生物学报. 2002, (03): 263-269
杨少丽,王印庚,董树刚.海水养殖鱼类弧菌病的研究进展[J].海洋水产研究. 2005, (04): 75-83
杨茂成.兽医统计学[M].北京:中国展望出版社, 1990: 232-234.
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