Involvement of PG2212 Zinc Finger Protein in the Regulation of Oxidative Stress Resistance in Porphyromonas gingivalis W83

Abstract

dThe adaptation of Porphyromonas gingivalis to H 2 O 2 -induced stress while inducible is modulated by an unknown OxyR-independent mechanism. Previously, we reported that the PG_2212 gene was highly upregulated in P. gingivalis under conditions of prolonged oxidative stress. Because this gene may have regulatory properties, its function in response to H 2 O 2 was further characterized. PG2212, annotated as a hypothetical protein of unknown function, is a 10.3-kDa protein with a cysteine 2-histidine 2 (Cys 2 His 2 ) zinc finger domain. The isogenic mutant P. gingivalis FLL366 (⌬PG_2212) showed increased sensitivity to H 2 O 2 and decreased gingipain activity compared to the parent strain. Transcriptome analysis of P. gingivalis FLL366 revealed that approximately 11% of the genome displayed altered expression (130 downregulated genes and 120 upregulated genes) in response to prolonged H 2 O 2 -induced stress. The majority of the modulated genes were hypothetical or of unknown function, although some are known to participate in oxidative stress resistance. The promoter region of several of the most highly modulated genes contained conserved motifs. In electrophoretic mobility shift assays, the purified rPG2212 protein did not bind its own promoter region but bound a similar region in several of the genes modulated in the PG_2212-deficient mutant. A metabolome analysis revealed that PG2212 can regulate a number of genes coding for proteins involved in metabolic pathways critical for its survival under the conditions of oxidative stress. Collectively, our data suggest that PG2212 is a transcriptional regulator that plays an important role in oxidative stress resistance and virulence regulation in P. gingivalis.

Differences in survival, virulence and biofilm formation between sialidase-deficient and W83 wild-type Porphyromonas gingivalis strains under stressful environmental conditions

“…McKenzie et al indicated that the inactivation of pg1372 (encoding a protein with DNA-binding properties) virtually eliminated the ability of P. gingivalis to adapt to oxidative stress [ 9 ]. Dou et al showed that a PG2212 (encoding a zinc finger protein) isogenic mutant strain of P. gingivalis exhibited an increased sensitivity to H 2 O 2 [ 10 ]. In this study, we cultured P. gingivalis W83 and its sialidase-deficient mutant strain under stressful environmental conditions, including various pH values, temperatures and oxidative stress conditions, and compared the growth, morphology, tolerance, virulence factors and biofilm formation of these two strains to investigate the roles of sialidase in the adaptation of P. gingivalis to these stressful conditions.…”

Studies of the extracytoplasmic function sigma factor PG0162 in Porphyromonas gingivalis

“…DNA microarray gene expression analyses were performed using the Roche NimbleGen customer arrays (100910_CW_P_ging_w83_expr_HX12) according to the standard NimbleGen procedure (NimbleGen Arrays User's Guide: Gene Expression Analysis v5.1) as previously reported previously (Dou et al ., ).…”

A Sialidase‐Deficient Porphyromonas gingivalis Mutant Strain Induces Less Interleukin‐1β and Tumor Necrosis Factor‐α in Epi4 Cells Than W83 Strain Through Regulation of c‐Jun N‐Terminal Kinase Pathway

“…Purified, recombined plasmid was used to transform △PG0352 via electroporation. Transformants were selected using TSB agar plates with 5 μg/mL clindamycin and 1 μg/mL tetracycline 25 …”

“…Transformants were selected using TSB agar plates with 5 mg/mL clindamycin and 1 mg/mL tetracycline. 25 Epi4 Cell Culture Epi4, an SV40 T-antigen-immortalized GEC line, 26 was maintained in medium ¶ supplemented with antibiotics (100 units/mL penicillin, 100 mg/mL streptomycin # ) at 37°C with 5% CO 2 .…”