Iron‐ and hemin‐dependent gene expression of Porphyromonas gingivalis

Although iron under anaerobic conditions is more accessible and highly reactive because of its reduced form, iron‐dependent regulation is not well known in anaerobic bacteria. Here, we investigated iron‐ and hemin‐dependent gene regulation in Porphyromonas gingivalis, an established periodontopathogen that primarily inhabits anaerobic pockets. Whole‐genome microarrays of P. gingivalis genes were used to compare the levels of gene expression under iron‐replete and iron‐depleted conditions as well as under hemin‐replete and hemin‐depleted conditions. Under iron‐depleted conditions, the expression of genes encoding proteins that participate in iron uptake and adhesion/invasion of host cells was increased, while that of genes encoding proteins involved in iron storage, energy metabolism, and electron transport was decreased. Interestingly, many of the genes with altered expression had no known function. Limiting the amount of hemin also resulted in a reduced expression of the genes encoding proteins involved in energy metabolism and electron transport. However, hemin also had a significant effect on many other biological processes such as oxidative stress protection and lipopolysaccharide synthesis. Overall, comparison of the data from iron‐depleted conditions to those from hemin‐depleted ones showed that although some regulation is through the iron derived from hemin, there also is significant distinct regulation through hemin only. Furthermore, our data showed that the molecular mechanisms of iron‐dependent regulation are novel as the deletion of the putative Fur protein had no effect on the expression of iron‐regulated genes. Finally, our functional studies demonstrated greater survivability of host cells in the presence of the iron‐stressed bacterium than the iron‐replete P. gingivalis cells. The major iron‐regulated proteins encoded by PG1019–20 may play a role in this process as deletion of these sequences also resulted in reduced survival of the bacterium when grown with eukaryotic cells. Taken together, the results of this study demonstrated the utility of whole‐genome microarray analysis for the identification of genes with altered expression profiles during varying growth conditions and provided a framework for the detailed analysis of the molecular mechanisms of iron and hemin acquisition, metabolism and virulence of P. gingivalis.