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2016 | 19 | 1 |

Tytuł artykułu

Modulation of virulence genes by the two-component system PhoP - PhoQ in avian pathogenic Escherichia coli

Treść / Zawartość

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Avian pathogenic Escherichia coli (APEC) infections are a very important problem in the poultry industry. PhoP-PhoQ is a two-component system that regulates virulence genes in APEC. In this study, we constructed strains that lacked the PhoP or PhoQ genes to assess regulation of APEC pathogenicity by the PhoP-PhoQ two-component system. The PhoP mutant strain AE18, PhoQ mutant strain AE19, and PhoP/PhoQ mutant strain AE20 were constructed by the Red homologous recombination method. Swim plates were used to evaluate the motility of the APEC strains, viable bacteria counting was used to assess adhesion and invasion of chick embryo fibroblasts, and Real-Time PCR was used to measure mRNA expression of virulence genes. We first confirmed that AE18, AE19, and AE20 were successfully constructed from the wild-type AE17 strain. AE18, AE19, and AE20 showed significant decreases in motility of 70.97%, 83.87%, and 37.1%, respectively, in comparison with AE17. Moreover, in comparison with AE17, AE18, AE19, and AE20 showed significant decreases of 63.11%, 65.42%, and 30.26%, respectively, in CEF cell adhesion, and significant decreases of 59.83%, 57.82%, and 37.90%, respectively, in CEF cell invasion. In comparison with AE17, transcript levels of sodA, polA, and iss were significantly decreased in AE18, while transcript levels of fimC and iss were significantly decreased in AE19. Our results demonstrate that deletion of PhoP or PhoQ inhibits invasion and adhesion of APEC to CEF cells and significantly reduces APEC virulence by regulating transcription of virulence genes.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

19

Numer

1

Opis fizyczny

p.31-40,fig.,ref.

Twórcy

autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
autor
  • College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China

Bibliografia

  • Bijlsma JJ, Groisman EA, et al. (2005) The PhoP/PhoQ system controls the intramacrophage type three secretion system of Salmonella enterica. Mol Microbiol 57: 85-96.
  • Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97: 6640-6645.
  • Garcia V, Escovi E, Soncini FC, Groisman EA, et al (1996) Mg + as an extracellular signal: environmental regulation of virulence. Cell 84: 165-174.
  • Kaper JB, Nataro JP, Mobley HL (2002) Pathogenic Escherichia coli. Nature Reviews Microbiology 2: 123-140.
  • Kasahara M, Atsuo Nakata, and Hideo Shinagawa (1992) Molecular Analysis of the Escherichia coli phoP-phoQ Operon. Bacteriol 174: 492-498.
  • Lertsethtakarn P, Ottemann KM, Hendrixson DR (2011) Motility and chemotaxis in Campylobacter and Helicobacter. Annual Review Microbiol 65: 389-410.
  • Ling H, Kang A, Tan MH, Qi X, Chang MW (2010) The absence of the luxS gene increases swimming motility and flagella synthesis in Escherichia coli K12. Biochem Biophys Res Commun 401: 521-526.
  • Muyrers J P, Zhang Y, Stewart A F (2001) Techniques Recombinogenic engineering-new options for cloning and manipulating DNA. Trends Biochem Sci 26:325-331.
  • Nam D, Choi E, Kweon DH, Shin D (2010) The RstB Sensor Acts on the PhoQ Sensor to Control Expression of PhoP-Regulated Genes. Molecules Cells 4: 363-368.
  • Ogasawara H, Shinohara S, Yamamoto, et al. (2012) Novel regulation targets of the metal-response BasS-BasR two-component system of Escherichia coli. Microbiol 158: 1482-1492.
  • Poteete AR (2001) What makes the bacteriophage lambdaRed system useful for genetic engineering: molecular mechanism and biological function. FEMS Microbiol Lett 201: 9-14.
  • Prost LR, Daley ME, Le Sage V, et al. (2007) Activation of the bacterial sensor kinase PhoQ by acidic pH. Mol Cell 26: 165-174.
  • Shen F, Hobley L, Doherty N, et al. (2010) In Helicobacter pylori auto-inducer-2 but not LuxS/MccAB catalysed reverse transsulphuration regulates motility through modulation of flagellar gene transcription. BMC Microbiol 10: 210.
  • Shprung T, Peleg A, Rosenfeld Y (2012) Effect of PhoPPhoQ activation by broad repertoire of antimicrobial peptides on bacterial resistance. J Biol Chem 7: 4544-4551.
  • Wang S, Niu C, Shi Z, Xia Y, Yaqoob M, Dai J, et al. (2011) Effects of ibeA mutant on virulence and biofilm formation of avian pathogenic Escherichia coli, Infect Immun 79: 279-287.
  • Yoshida A, Ansai T, Takehara T, Kuramitsu HK (2005) LuxS-based signaling affects Streptococcus mutans biofilm formation. Appl Environ Microbiol 71: 2372-2380.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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