期刊论文详细信息
BMC Microbiology
TypA is involved in virulence, antimicrobial resistance and biofilm formation in Pseudomonas aeruginosa
Joerg Overhage2  Olivier Lesouhaitier3  Martina Rueger2  Nikola Strempel2  Beatrix Tettmann2  Thibaut Rosay3  Amy TY Yeung1  Anke Neidig2 
[1] Centre for Microbial Diseases & Immunity Research, University of British Columbia, 2259 Lower Mall, Vancouver, BC, Canada;Karlsruhe Institute of Technology (KIT), Institute of Functional Interfaces, PO Box 3640, Karlsruhe, 76021, Germany;Laboratory of Microbiology Signals and Microenvironment, LMSM EA 4312, University of Rouen, 55 rue Saint Germain, Evreux, 27000, France
关键词: Resistance;    Biofilm;    Macrophage;    Dictyostelium discoideum;    Virulence;    Type III secretion system;    TypA;    Pathogen;    Pseudomonas aeruginosa;   
Others  :  1143955
DOI  :  10.1186/1471-2180-13-77
 received in 2012-10-30, accepted in 2013-04-04,  发布年份 2013
PDF
【 摘 要 】

Background

Pseudomonas aeruginosa is an important opportunistic human pathogen and is extremely difficult to treat due to its high intrinsic and adaptive antibiotic resistance, ability to form biofilms in chronic infections and broad arsenal of virulence factors, which are finely regulated. TypA is a GTPase that has recently been identified to modulate virulence in enteric Gram-negative pathogens.

Results

Here, we demonstrate that mutation of typA in P. aeruginosa resulted in reduced virulence in phagocytic amoebae and human macrophage models of infection. In addition, the typA mutant was attenuated in rapid cell attachment to surfaces and biofilm formation, and exhibited reduced antibiotic resistance to ß-lactam, tetracycline and antimicrobial peptide antibiotics. Quantitative RT-PCR revealed the down-regulation, in a typA mutant, of important virulence-related genes such as those involved in regulation and assembly of the Type III secretion system, consistent with the observed phenotypes and role in virulence of P. aeruginosa.

Conclusions

These data suggest that TypA is a newly identified modulator of pathogenesis in P. aeruginosa and is involved in multiple virulence-related characteristics.

【 授权许可】

   
2013 Neidig et al.; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20150330045858455.pdf 453KB PDF download
Figure 4. 38KB Image download
Figure 3. 42KB Image download
Figure 2. 28KB Image download
Figure 1. 21KB Image download
【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】
  • [1]Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FS, Hufnagle WO, Kowalik DJ, Lagrou M: Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 2000, 406(6799):959-964.
  • [2]Govan JR, Deretic V: Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 1996, 60(3):539-574.
  • [3]Breidenstein EB, de la Fuente-Nunez C, Hancock RE: Pseudomonas aeruginosa: all roads lead to resistance. Trends Microbiol 2011, 19(8):419-426.
  • [4]Feinbaum RL, Urbach JM, Liberati NT, Djonovic S, Adonizio A, Carvunis AR, Ausubel FM: Genome-wide identification of pseudomonas aeruginosa virulence-related genes using a caenorhabditis elegans infection model. PLoS Pathog 2012, 8(7):e1002813.
  • [5]Hauser AR: The type III secretion system of Pseudomonas aeruginosa: infection by injection. Nat Rev Microbiol 2009, 7(9):654-665.
  • [6]Filloux A: Protein secretion systems in pseudomonas aeruginosa: an essay on diversity, evolution, and function. Front Microbiol 2011, 2:155.
  • [7]Girard G, Bloemberg GV: Central role of quorum sensing in regulating the production of pathogenicity factors in Pseudomonas aeruginosa. Future Microbiol 2008, 3(1):97-106.
  • [8]Smith RS, Iglewski BH: P. aeruginosa quorum-sensing systems and virulence. Curr Opin Microbiol 2003, 6(1):56-60.
  • [9]Aballay A, Ausubel FM: Caenorhabditis elegans as a host for the study of host-pathogen interactions. Curr Opin Microbiol 2002, 5(1):97-101.
  • [10]Lima WC, Lelong E, Cosson P: What can Dictyostelium bring to the study of Pseudomonas infections? Semin Cell Dev Biol 2011, 22(1):77-81.
  • [11]Limmer S, Quintin J, Hetru C, Ferrandon D: Virulence on the fly: drosophila melanogaster as a model genetic organism to decipher host-pathogen interactions. Curr Drug Targets 2011, 12(7):978-999.
  • [12]Wang F, Zhong NQ, Gao P, Wang GL, Wang HY, Xia GX: SsTypA1, a chloroplast-specific TypA/BipA-type GTPase from the halophytic plant Suaeda salsa, plays a role in oxidative stress tolerance. Plant Cell Environ 2008, 31(7):982-994.
  • [13]Scott K, Diggle MA, Clarke SC: TypA is a virulence regulator and is present in many pathogenic bacteria. Br J Biomed Sci 2003, 60(3):168-170.
  • [14]Verstraeten N, Fauvart M, Versees W, Michiels J: The universally conserved prokaryotic GTPases. Microbiol Mol Biol Rev 2011, 75(3):507-542. second and third pages of table of contents
  • [15]DeLivron MA, Robinson VL: Salmonella enterica serovar Typhimurium BipA exhibits two distinct ribosome binding modes. J Bacteriol 2008, 190(17):5944-5952.
  • [16]Britton RA: Role of GTPases in bacterial ribosome assembly. Annu Rev Microbiol 2009, 63:155-176.
  • [17]Hwang J, Tseitin V, Ramnarayan K, Shenderovich MD, Inouye M: Structure-based design and screening of inhibitors for an essential bacterial GTPase, Der. J Antibiot (Tokyo) 2012, 65(5):237-243.
  • [18]Grant AJ, Farris M, Alefounder P, Williams PH, Woodward MJ, O’Connor CD: Co-ordination of pathogenicity island expression by the BipA GTPase in enteropathogenic Escherichia coli (EPEC). Mol Microbiol 2003, 48(2):507-521.
  • [19]Farris M, Grant A, Richardson TB, O’Connor CD: BipA: a tyrosine-phosphorylated GTPase that mediates interactions between enteropathogenic Escherichia coli (EPEC) and epithelial cells. Mol Microbiol 1998, 28(2):265-279.
  • [20]Kiss E, Huguet T, Poinsot V, Batut J: The typA gene is required for stress adaptation as well as for symbiosis of Sinorhizobium meliloti 1021 with certain Medicago truncatula lines. Mol Plant Microbe Interact 2004, 17(3):235-244.
  • [21]Beckering CL, Steil L, Weber MH, Volker U, Marahiel MA: Genomewide transcriptional analysis of the cold shock response in Bacillus subtilis. J Bacteriol 2002, 184(22):6395-6402.
  • [22]Overhage J, Lewenza S, Marr AK, Hancock RE: Identification of genes involved in swarming motility using a Pseudomonas aeruginosa PAO1 mini-Tn5-lux mutant library. J Bacteriol 2007, 189(5):2164-2169.
  • [23]Breidenstein EBM L, Janot J, Strehmel L, Fernandez PK, Taylor I, Kukavica-Ibrulj SL, Gellatly RC, Levesque J, Overhage LJ, Hancock REW: The Lon protease is essential for full virulence in Pseudomonas aeruginosa. PLoS One 2012, 7(11):e49123.
  • [24]Adamek M, Overhage J, Bathe S, Winter J, Fischer R, Schwartz T: Genotyping of environmental and clinical Stenotrophomonas maltophilia isolates and their pathogenic potential. PLoS One 2011, 6(11):e27615.
  • [25]Liberati NT, Urbach JM, Miyata S, Lee DG, Drenkard E, Wu G, Villanueva J, Wei T, Ausubel FM: An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants (vol 103, pg 2833, 2006). P Natl Acad Sci USA 2006, 103(52):19931-19931.
  • [26]Saliba AM, Filloux A, Ball G, Silva ASV, Assis MC, Plotkowski MC: Type III secretion-mediated killing of endothelial cells by Pseudomonas aeruginosa. Microb Pathogenesis 2002, 33(4):153-166.
  • [27]Tan MW, Rahme LG, Sternberg JA, Tompkins RG, Ausubel FM: Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P. aeruginosa virulence factors. P Natl Acad Sci USA 1999, 96(5):2408-2413.
  • [28]Duo M, Hou S, Ren D: Identifying Escherichia coli genes involved in intrinsic multidrug resistance. Appl Microbiol Biotechnol 2008, 81(4):731-741.
  • [29]Matz C, Moreno AM, Alhede M, Manefield M, Hauser AR, Givskov M, Kjelleberg S: Pseudomonas aeruginosa uses type III secretion system to kill biofilm-associated amoebae. ISME J 2008, 2(8):843-852.
  • [30]Aiello D, Williams JD, Majgier-Baranowska H, Patel I, Peet NP, Huang J, Lory S, Bowlin TL, Moir DT: Discovery and characterization of inhibitors of Pseudomonas aeruginosa type III secretion. Antimicrob Agents Chemother 2010, 54(5):1988-1999.
  • [31]DeLivron MA, Makanji HS, Lane MC, Robinson VL: A novel domain in translational GTPase BipA mediates interaction with the 70S ribosome and influences GTP hydrolysis. Biochemistry 2009, 48(44):10533-10541.
  • [32]Sircili MP, Walters M, Trabulsi LR, Sperandio V: Modulation of enteropathogenic Escherichia coli virulence by quorum sensing. Infect Immun 2004, 72(4):2329-2337.
  • [33]Micklinghoff JC, Schmidt M, Geffers R, Tegge W, Bange FC: Analysis of expression and regulatory functions of the ribosome-binding protein TypA in Mycobacterium tuberculosis under stress conditions. Arch Microbiol 2010, 192(6):499-504.
  • [34]Yahr TL, Wolfgang MC: Transcriptional regulation of the Pseudomonas aeruginosa type III secretion system. Mol Microbiol 2006, 62(3):631-640.
  • [35]Wareham DW, Papakonstantinopoulou A, Curtis MA: The Pseudomonas aeruginosa PA14 type III secretion system is expressed but not essential to virulence in the Caenorhabditis elegans-P. aeruginosa pathogenicity model. FEMS Microbiol Lett 2005, 242(2):209-216.
  • [36]Darby C, Cosma CL, Thomas JH, Manoil C: Lethal paralysis of Caenorhabditis elegans by Pseudomonas aeruginosa. P Natl Acad Sci USA 1999, 96(26):15202-15207.
  • [37]Qi SY, Li Y, Szyroki A, Giles IG, Moir A, O’Connor CD: Salmonella typhimurium responses to a bactericidal protein from human neutrophils. Mol Microbiol 1995, 17(3):523-531.
  • [38]Barker HC, Kinsella N, Jaspe A, Friedrich T, O’Connor CD: Formate protects stationary-phase Escherichia coli and Salmonella cells from killing by a cationic antimicrobial peptide. Mol Microbiol 2000, 35(6):1518-1529.
  • [39]Hoiby N, Ciofu O, Johansen HK, Song ZJ, Moser C, Jensen PO, Molin S, Givskov M, Tolker-Nielsen T, Bjarnsholt T: The clinical impact of bacterial biofilms. Int J Oral Sci 2011, 3(2):55-65.
  • [40]Jensen PO, Givskov M, Bjarnsholt T, Moser C: The immune system vs Pseudomonas aeruginosa biofilms. FEMS Immunol Med Microbiol 2010, 59(3):292-305.
  • [41]Mah TF, O’Toole GA: Mechanisms of biofilm resistance to antimicrobial agents. Trends Microbiol 2001, 9(1):34-39.
  • [42]West SE, Schweizer HP, Dall C, Sample AK, Runyen-Janecky LJ: Construction of improved Escherichia-Pseudomonas shuttle vectors derived from pUC18/19 and sequence of the region required for their replication in Pseudomonas aeruginosa. Gene 1994, 148(1):81-86.
  • [43]Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP: A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 1998, 212(1):77-86.
  • [44]Yeung AT, Bains M, Hancock RE: The sensor kinase CbrA is a global regulator that modulates metabolism, virulence, and antibiotic resistance in Pseudomonas aeruginosa. J Bacteriol 2011, 193(4):918-931.
  • [45]Fey P, Kowal AS, Gaudet P, Pilcher KE, Chisholm RL: Protocols for growth and development of Dictyostelium discoideum. Nat Protoc 2007, 2(6):1307-1316.
  • [46]Amiel E, Acker JL, Collins RM, Berwin B: Uncoupling scavenger receptor A-mediated phagocytosis of bacteria from endotoxic shock resistance. Infect Immun 2009, 77(10):4567-4573.
  • [47]Sulston J, Hodgkin J: The Nematode Caenorhabditis elegans. Wood: W. B; 1988.
  • [48]Stiernagle T: Maintenance of C. elegans. In C. elegans. A practical approach. Edited by Hope IA. Oxford, United Kingdom: Oxford University Press; 1999:51-67.
  • [49]Blier AS, Veron W, Bazire A, Gerault E, Taupin L, Vieillard J, Rehel K, Dufour A, Le Derf F, Orange N: C-type natriuretic peptide modulates quorum sensing molecule and toxin production in Pseudomonas aeruginosa. Microbiology 2011, 157(Pt 7):1929-1944.
  • [50]Wiegand I, Hilpert K, Hancock RE: Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc 2008, 3(2):163-175.
  • [51]Friedman L, Kolter R: Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol Microbiol 2004, 51(3):675-690.
  • [52]Marr AK, Overhage J, Bains M, Hancock RE: The Lon protease of Pseudomonas aeruginosa is induced by aminoglycosides and is involved in biofilm formation and motility. Microbiology 2007, 153(Pt 2):474-482.
  文献评价指标  
  下载次数:8次 浏览次数:11次