55 ± 0.18 sitA 2.81 ± 0.08 selleck chemicals a Mean expression ratio (±SD) of ΔfurΔryhB relative to Δfur. Discussion In this study, we provide an initial characterisation of K. pneumoniae RyhB. In K. pneumoniae, sequence comparison indicated that the
nucleotide sequence of the ryhB gene (91 bp) is 92.3% identical to the E. coli version (90 bp). However, the promoter sequence of K. pneumoniae ryhB is only 72.4% identical to that of E. coli. In this study, we found that the expression of ryhB in K. pneumoniae is directly repressed by Fur-Fe(II), as is the case in E. coli (Figure 1). In addition, structure of the genomic neighbourhood of ryhB differs between the 2 species. In the E. coli genome, ryhB is found between yhhX and yhhY. In the K. pneumoniae genome, ryhB is flanked by yhhY and a hypothetical ORF. By Pfam search, the hypothetical ORF was found to contain a bactofilin domain (E-value = 3.7e-24), which belongs to a new class of polymer-forming proteins that serve as versatile molecular scaffolds in a selleck chemicals llc variety of cellular pathways . Even though the function of this hypothetical protein in K. pneumoniae has not yet been investigated, we found that RyhB could strongly repress the expression of this hypothetical protein (unpublished data). This result PF-02341066 order suggests that RyhB could participate in a variety of cellular pathways in K. pneumoniae. We previously showed in K. pneumoniae, Fur represses CPS biosynthesis via regulation
of RmpA, RmpA2, and RcsA. In addition to these 3 regulators, Enzalutamide research buy one or more regulators may be involved in the Fur-mediated control of cps transcription . In this study, we found that RyhB also participates in Fur-regulated CPS biosynthesis
via activation of orf1 and orf16 transcription and is independent of the 3 regulators, RmpA, RmpA2, and RcsA (Figure 2 and 3). We want to further analyse whether any potential transcriptional regulator-binding motifs exist in the promoter sequences of orf1 and orf16. We noted that a binding site typical of IscR, a transcriptional repressor that controls Fe–S biosynthesis , was located 172 bp upstream of the translation start site of GalF (encoded by orf1, 5′-ATAACCTGAACGAAAATAAGATTAT-3′). The predication indicated that IscR could participate in control of orf1 expression. Furthermore, a previous study reported that RyhB promotes the degradation of iscSUA transcripts, resulting in an increase in the ratio of apo-IscR/holo-IscR . Whether RyhB activates CPS biosynthesis via regulation of the ratio of apo-IscR/holo-IscR in K. pneumoniae awaits further analysis. However, the regulatory mechanism of cps transcription is more complex than expected; whether another unknown transcriptional regulator is involved in activation of RyhB’s effect on orf16 transcription needs to be investigated. In addition, CPS is considered the major determinant that can protect the bacteria from phagocytosis and killing by serum factors [8, 9].