The first gene (HI1010) is a potential 6-phosphogluconate dehydro

The first gene (HI1010) is a potential 6-phosphogluconate dehydrogenase that generates ribulose-5-phosphate. This links directly into the PPP and other energy and biosynthetic pathways (outlined in Figure 3). Table 2 Genes

differentially expressed in H. influenzae Eagan at pH 8.0 compared to pH 6.8 Genes up-regulated at pH 8.0 compared to 6.8 Metabolic genes Gene Log 2 fold p -value FDR Comment HI1010 2.21 5.12×10-10 1.02×10-7 6-phosphogluconate dehydrogenase HI1011 2.20 6.83×10-10 1.22×10-7 Similar to YgbK HI1012 2.04 3.06×10-8 3.BLZ945 64×10-6 Sugar isomerase HI1013 1.88 3.04×10-7 2.86×10-5 Hydroxypyruvate isomerase HI1014 1.52 2.33×10-5 1.54×10-3 Sugar epimerase HI1015 1.12 1.18×10-3 AC220 molecular weight 4.70×10-2 GntP family, gluconate:H+ symporter HI0091 1.74 5.98×10-7 5.33×10-5 Hypothetical protein; homologous to GlxK, glycerate kinase HI0092 2.14 1.49×10-9 2.41×10-7 GntP family, gluconate:H+ symporter Iron uptake genes Gene Log 2 fold p -value FDR Comment HI0995 1.53 1.72×10-5 1.23×10-3 OMP, iron-binding hitA 2.21 1.69×10-10 3.77×10-8 Iron uptake hxuB 1.65 1.54×10-6 1.25×10-4 Hemopexin utilization protein hxuC 1.70 8.04×10-7 6.83×10-5 TonB-dependent heme receptor Genes of unknown function Gene Log 2 fold p -value FDR Comment HI1427 1.54 6.87×10-6 5.33×10-4 Hypothetical protein Genes down-regulated at pH 8.0 compared to 6.8 Gene Log 2 fold p -value FDR Comment HI1349 -2.31 5.58×10-11 1.42×10-8 Ferritin

HI1385 -1.55 2.27×10-5 1.54×10-3 FtnB; non-heme ferritin Figure 3 The pathway uniquely induced in H. influenzae Eagan at pH 8.0. (A) Genes HI1010-1015 (block arrows, grey) were all induced in Nirogacestat in vitro H. influenzae Eagan at pH 8.0. In silico analysis identified 2 promoters

across this region of the genome (indicated by line arrows) and HI1010-HI1015 forms a single operon. (B) These HI1010-1015 genes encode a gluconate:H+ symporter, a putative 6-phospohogluconate dehydrogenase and a range of sugar isomerases and epimerases that would link gluconate to the PPP and other metabolic pathways (the putative role for these genes are shown in blue). The GntP symporter family of transporters also import H+, as part of the survival response associated with an increased environmental pH (Table 2). It is interesting Tenofovir supplier to note that our bioinformatic analyses have identified an operator/promoter upstream of HI1010 (Figure 3) with a putative DeoR binding site; HI1010 is divergent to a DeoR-like gene. While not within the scope of this project it is known in other bacteria that DeoR-like regulators variously control pathways directing sugar metabolism and are connected to the PPP. Also, the bioinformatics analyses indicate that the HI1010-1015 genes are on a single transcriptional unit, forming an operon. Traditionally high concentrations of glucose are thought to be oxidized extracellularly by membrane-bound dehydrogenases.

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