Table 1.
List of strains and plasmids used in this study.
Table 2.
List of primers used in this study.
Table 3.
The relative expression of prtA, psaB, psaC, psaA, and pcpA genes was normalized with the housekeeping gene gyrA.
The log2 fold increase is relative to the expression in the D39 wild-type grown in CDMchelex with 0.3 mM Ni2+ to that with 0 mM Ni2+. Standard deviation of three independent replications is given in parentheses.
Table 4.
β-galactosidase activity (miller units) of PpcpA-lacZ, PpsaB-lacZ, and PprtA-lacZ in S. pneumoniae D39 wild-type and ΔpsaR (RW100)grown in CDMchelex and CDMchelex-Mn2+ supplemented with various concentrations of Ni2+ (mM).
Standard deviation of three independent replications is given in parentheses, whereas ND stands for not determined. Noteworthy, lacZ was fused to the 3’ end of prtA* on the native chromosomal location, using plasmid pOR113. This might explain the lower Miller Units of PprtA compared to PpcpA and PpsaB.
Table 5.
Summary of transcriptome comparison of S. pneumoniae D39 wild-type strain with ΔpsaR grown in CDM with 0.3 mM Ni2+.
Table 6.
Expression level (in Miller units) of PpcpA-lacZ, PpsaB-lacZ, and PprtA-lacZ in D39 wild-type in CDMchelex and CDMchelex-Mn2+ supplemented with different concentrations of Ni2+ and Mn2+ (mM).
Standard deviation of three independent replicates is indicated in bars.
Fig 1.
In vitro interaction of PsaR-Strep tag with the promoter regions of pcpA (A), psaB (B), prtA (C), and phtB (D).
PsaR-Strep was added at concentration of 30 nM as indicated above panel, while lane 1 is without added protein. Arrows indicate the position of shifted probe and asterisks indicate the position of free probe. Mn2+ was added with concentrations of 0.05 mM in lanes 3, 7, and 9, and 0.1 mM in lane 4, 8, and 10. Ni2+ was added with concentrations of 0.2 mM in lanes 5, 7, and 9, and 0.4 mM in lanes 6, 8, and 10.
Fig 2.
(A) Cell-associated metal ion concentrations (expressed ug g-1) of S. pneumoniae D39 wild type when grown in CDMchelex with either 0 mM or 0.3 mM Ni2+. (B) Metal ions contents of S. pneumoniae D39 wild-type when grown in CDMchelex containing 0.02 mM Mn2+ with addition of 0, 0.1, 0.3 or 0.5 mM Ni2+. The statistical significance of the differences in the mean metal concentrations was determined by one-way ANOVA (NS not significant, *P<0.01, and ***P<0.0001)