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Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

Departments of Pharmacology,¹ Oral Infectious Diseases and Immunology, Graduate School of Dental Science, Kyushu University,⁴ Special Patient Oral Care Unit, Kyushu University Dental Hospital, Fukuoka 812-8582,² Division of Microbiology and Oral Infection, Course of Medical and Dental Sciences, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588,³ Department of Microbiology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan⁵

Received 29 April 2002/ Returned for modification 14 August 2002/ Accepted 18 November 2002

The periodontopathogen Porphyromonas gingivalis is an obligate anaerobe that is devoid of catalase but exhibits a relatively high degree of resistance to peroxide stress. In the present study, we demonstrate that P. gingivalis contains a Dps homologue that plays an important role in the protection of cells from peroxide stress. The Dps protein isolated from P. gingivalis displayed a ferritin-like spherical polymer consisting of 19-kDa subunits. Molecular cloning and sequencing of the gene encoding this protein revealed that it had a high similarity in nucleotide and amino acid sequences to Dps proteins from other species. The expression of Dps was significantly increased by exposure of P. gingivalis to atmospheric oxygen in an OxyR-dependent manner, indicating that it is regulated by the reactive oxygen species-regulating gene oxyR. The Dps-deficient mutants, including the dps single mutant and the ftn dps double mutant, showed no viability loss upon exposure to atmospheric oxygen for 6 h. In contrast to the wild type, however, these mutants exhibited the high susceptibility to hydrogen peroxide, thereby disrupting the viability. On the other hand, no significant difference in sensitivity to mitomycin C and metronidazole was observed between the wild type and the mutants. Furthermore, the dps single mutant, compared with the wild type, showed a lower viability in infected human umbilical vein endothelial cells.

Editor: V. J. DiRita

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