Identification of an HptB-mediated multi-step phosphorelay in Pseudomonas aeruginosa PAO1

PecS regulates the urate-responsive expression of type 1 fimbriae in Klebsiella pneumoniae CG43

“…The putative promoter regions of pecS, pecM, fimA and fimB were PCR-amplified using primers (Lin et al, 2006) to generate P pecS -lacZ, P pecM -lacZ, P fimA -lacZ and P fimB -lacZ. The promoter-reporter plasmids were individually mobilized into K. pneumoniae CG43S3DlacZ strains through conjugation from E. coli S17-1 lpir.…”

“…Each sample was assayed in triplicate, and at least three independent experiments were conducted. The data were calculated from three independent experiments and are shown as the mean+SD from the nine samples (Lin et al, 2006). The pH of the modified LB broth, which had been supplemented with different concentrations of urate (dissolved in 1 M NaOH), was adjusted to 7.5 with HCl as described in Wilkinson & Grove (2004).…”

FimK regulation on the expression of type 1 fimbriae in Klebsiella pneumoniae CG43S3

“…As described previously (Lin et al, 2006;Wu et al, 2010), bacteria diluted 1 : 100 in LB broth supplemented with appropriate antibiotics were inoculated into each well of a 96-well microtitre dish (Orange Scientific) and statically incubated at 37 uC for 20 h. After removing planktonic cells, the wells were washed once with distilled water to remove unattached cells. Crystal violet (0.1 % w/v; Sigma-Aldrich) was used to stain the attached cells for 30 min.…”

“…The putative promoter regions of fimA, fimB and fimE were PCR-amplified with primers pfimA3/ pfimA4, pfimB4/pfimB5 and pfimE1/pfimE2, respectively. The amplicons were then cloned into placZ15 (Lin et al, 2006) to generate P fimA -lacZ (locked on), P fimB -lacZ and P fimE -lacZ. The promoter-reporter plasmids were individually mobilized into K. pneumoniae CG43S3 strains by conjugation from E. coli S17-1lpir.…”

Sticky Situations: Key Components That Control Bacterial Surface Attachment

“…HptB is proposed to regulate rsmY levels in a GacA-dependent manner via as-yet-unknown factors (16). This, however, is not true of SagS, a sensor-regulator hybrid that participates in a phosphotransfer event with HptB (63,95) and is involved in the regulation of attachment and biofilm formation (129). SagS suppresses sRNA levels predominantly under planktonic growth conditions, with inactivation of sagS resulting in a temporary enhancement of attachment but a defect in the later stages of biofilm formation (129).…”