title: Nitric oxide stress resistance in Porphyromonas gingivalis is mediated by a putative hydroxylamine reductase.
abstract: Porphyromonas gingivalis, the causative agent of adult periodontitis, must maintain nitric oxide (NO) homeostasis and surmount nitric oxide stress from host immune responses or other oral bacteria to survive in the periodontal pocket. To determine the involvement of a putative hydroxylamine reductase (PG0893) and a putative nitrite reductase-related protein (PG2213) in P. gingivalis W83 NO stress resistance, genes encoding those proteins were inactivated by allelic exchange mutagenesis. The isogenic mutants P. gingivalis FLL455 (PG0893ermF) and FLL456 (PG2213ermF) were black pigmented and showed growth rates and gingipain and hemolytic activities similar to those of the wild-type strain. P. gingivalis FLL455 was more sensitive to NO than the wild type. Complementation of P. gingivalis FLL455 with the wild-type gene restored the level of NO sensitivity to a level similar to that of the parent strain. P. gingivalis FLL455 and FLL456 showed sensitivity to oxidative stress similar to that of the wild-type strain. DNA microarray analysis showed that PG0893 and PG2213 were upregulated 1.4- and 2-fold, respectively, in cells exposed to NO. In addition, 178 genes were upregulated and 201 genes downregulated more than 2-fold. The majority of these modulated genes were hypothetical or of unknown function. PG1181, predicted to encode a transcriptional regulator, was upregulated 76-fold. Transcriptome in silico analysis of the microarray data showed major metabolomic variations in key pathways. Collectively, these findings indicate that PG0893 and several other genes may play an important role in P. gingivalis NO stress resistance.
authors.affiliation: Division of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, Loma Linda, California, USA. mboutrin@llu.edu
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Boutrin
authors.forename: Charles
authors.initials: C
authors.lastname: Wang
authors.forename: Wilson
authors.initials: W
authors.lastname: Aruni
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Li
authors.forename: Hansel M
authors.initials: HM
authors.lastname: Fletcher
mesh_terms: D024881:Drug Resistance, Bacterial; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D046228:Microarray Analysis; D009569:Nitric Oxide / Q000633:toxicity*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D016966:Porphyromonas gingivalis / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D013312:Stress, Physiological
chemical_list: D009569:Nitric Oxide; D010088:Oxidoreductases; C021402:hydroxylamine reductase