Table 1 Crystal sizes in various strains under different conditions Strain Anaerobic nitrate medium Microaerobic nitrate medium WT 38.0 ± 15.8 nm 30.5 ± 12.4 nm ΔMgfnr mutant 40.2 ± 15.3 nm 21.9 ± 7.7 nm WT + pLYJ110 Midostaurin in vivo 30.3 ± 15.1 nm 23.5 ± 13.8 nm ΔMgfnr + pLYJ110 42.1 ± 21.9 nm 30.3 ± 22.3 nm WT + pLYJ153 31.7 ± 18.7 nm 30.0 ± 21.6 nm ΔMgfnr + pLYJ153 40.9 ± 20.2 nm 31.3 ± 20.7 nm In ΔMgfnr expression patterns of denitrification genes are different from those in WT Deletion of Mgfnr resulted in impaired magnetite biomineralization only under microaerobic conditions
in the presence of nitrate, suggesting a potential link to nitrate reduction. In addition, in E. coli and other bacteria, Fnr was shown to upregulate the expression of denitrification genes under microaerobic or anaerobic conditions [30, 31]. Our earlier studies learn more on MSR-1 showed that a complete denitrification pathway including genes encoding
for nitrate (nap), nitrite (nir), nitric oxide (nor), and nitrous oxide reduction (nos) occurs for anaerobic growth. In addition, all denitrification genes in the WT were regulated by oxygen, and except for nap, which was upregulated by oxygen, the highest expression of other denitrification genes coincided with conditions permitting maximum magnetosome formation (e.g., low oxygen tensions and the
presence of nitrate) [5]. Consistent with this, we found putative Fnr binding sites (TTGA N 6 TCAA) in the promoter regions of all operons involved in denitrification (Additional file 2). To gain insight whether these observed defects in magnetosome formation in ΔMgfnr strain are indirectly caused by deregulation of denitrification genes, we analyzed the transcription of all denitrification genes by constructing gusA fusions in the ΔMgfnr background (Table 2). In ΔMgfnr strain, expression of nap was no longer upregulated by oxygen but displayed similar levels of β-glucuronidase activity under all tested conditions, which was higher than the maximum level in the WT. nirS-gusA showed a similar Bay 11-7085 pattern as in WT, that is, it was upregulated by nitrate and downregulated by oxygen. However, an about 5-fold higher β-glucuronidase activity was measured under aerobic conditions compared to the WT. ΔMgfnr mutant cells harboring the transcriptional nor-gusA reporter gene fusion exhibited a higher β-glucuronidase activity under microaerobic conditions in the presence of nitrate (416 U/mg) than in the absence of nitrate (151 U/mg), while it was lower than in the WT under the same conditions. However, oxygen did not inhibit the expression of nor-gusA in the ΔMgfnr strain.