Defining the role of Hmu and Hus systems in Porphyromonas gingivalis heme and iron homeostasis and virulence

Iron and heme are essential nutrients for all branches of life. Pathogenic members of the Bacteroidota phylum, including Porphyromonas gingivalis, do not synthesize heme and rely on host hemoproteins for heme as a source of iron and protoporphyrin IX. P. gingivalis is the main pathogen responsible for dysbiosis in the oral microbiome and the initiation and progression of chronic periodontitis. It utilizes Hmu and Hus systems for heme uptake, including HmuY and HusA hemophore-like proteins and their cognate HmuR and HusB TonB-dependent outer membrane heme receptors. Although the mechanisms of heme uptake are relatively well characterized in P. gingivalis, little is known about the importance of heme uptake systems in heme and iron homeostasis and virulence. Therefore, this work aimed to investigate these mechanisms in detail. We characterized the P. gingivalis double mutant strain deficient in functional hmuY and hmuR or husA and husB genes. Global gene expression and phenotypic analyses revealed that the Hmu system significantly influences heme homeostasis, confirming its main role in heme supply. Both systems, particularly the Hus system, affect the virulence of P. gingivalis. Our results demonstrate the diverse role of Hmu and Hus systems in P. gingivalis heme and iron homeostasis and virulence.