title: Polyphosphate kinase 1 is required for the pathogenesis process of meningitic Escherichia coli K1 (RS218).
abstract: OBJECTIVE
  Polyphosphate kinase 1 (PPK1), encoded by the ppk1 gene, is one of the major enzymes to reversibly catalyze the synthesis of polyphosphate (poly P) from the terminal phosphate of ATP. Poly P confers resistance to stress in a number of bacterial species but its role in the virulence of meningitic bacterial pathogens is unknown. The aim of this study was to determine the role of PPK1 in the pathogenesis of Escherichia coli meningitis.
  
  
  METHODS
  An isogenic in-frame ppk1 deletion mutant (PD44) of E. coli K1 strain E44 was constructed and characterized. Human brain microvascular endothelial cells and neonatal rats were used as the in vitro and in vivo models, respectively, to evaluate bacterial adhesion/invasion and the abilities of bacteria crossing the blood-brain barrier (BBB) to cause meningitis. The survival of PD44 and E44 under osmotic and acid stress conditions were also examined.
  
  
  RESULTS
  Poly P levels in E44 were clearly higher than those in PD44, especially at the stationary phase (SP). The ppk1 deletion mutant PD44 also showed poor survival rates during osmotic shock and acidic challenge, which the bacteria would face during pathogenesis. In vitro and in vivo assays revealed that PD44 was defective in bacterial adhesion and translocation across the BBB. By using the Evans blue method, we found that E44-induced permeability of the BBB in neonatal rats was significantly higher than that of the animals infected with PD44. Cytokine ELISA results showed that the TNF-α and IL-1β levels in the serum and brain tissues of the neonatal rats infected with PD44 were lower than that of the E44 group. A more obvious meningeal inflammation could be observed in the brain tissues of the rats infected with E44 when compared with that of the PD44 group by histopathological examination. Furthermore, the mRNA expression of IbeR, which is an RpoS-like regulator contributing to the SP regulation in E44, was found to be decreased in PD44 when compared with the parent strain. PD44 was also deficient in mRNA expression of the invasin IbeA, the adhesin FimH and the outer member protein A, which contributes to E44 penetration across BBB and resistance to the stimulations of low pH and high osmolarity.
  
  
  CONCLUSIONS
  These results indicate that ppk1 plays an important role in stress adaption and virulence in meningitic E. coli K1 strain E44, and controls the relevant phenotypes by modulating the expression of the SP regulatory gene ibeR and the virulence genes ibeA, fimH and ompA.
authors.affiliation: Department of Microbiology, School of Public Health & Tropical Medicine, Southern Medical University, Tonghe, Guangzhou, 510515, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Peng
authors.forename: Wen-Ying
authors.initials: WY
authors.lastname: Luo
authors.forename: Tie
authors.initials: T
authors.lastname: Zhao
authors.forename: Cheng-Song
authors.initials: CS
authors.lastname: Wan
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
authors.forename: Feng
authors.initials: F
authors.lastname: Chi
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.forename: Hong
authors.initials: H
authors.lastname: Cao
authors.forename: Sheng-He
authors.initials: SH
authors.lastname: Huang
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008297:Male; D020814:Meningitis, Escherichia coli / Q000382:microbiology*; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D011122:Polyphosphates / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014774:Virulence
chemical_list: D029968:Escherichia coli Proteins; D011122:Polyphosphates; D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase