PG1659 functions as anti‐sigma factor to extracytoplasmic function sigma factor RpoE in <i>Porphyromonas gingivalis</i> W83

Dou, Youjun; Rutanhira, Hiel; Schormann, Norbert; Deivanayagam, Champion; Fletcher, Hansel M.

doi:10.1111/omi.12329

Cited by 5 publications

(1 citation statement)

References 63 publications

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“…The Rgp and Kgp protease activities were determined using BAPNA (Nα‐benzoyl‐ dl ‐arginine‐ p ‐nitroanilide; 4 mM) and ALNA (AC‐Lys‐ p ‐nitroanilide HCl; 4 mM) as substrates in an activated protease buffer (0.1 M Tris, pH = 7.6, 0.2 M NaCl, 5 mM CaCl 2 , and 9 mM l ‐cysteine) (Dou et al., 2021). The P. gingivalis culture (OD = 1.5) was centrifuged at 6000 × g for 10 min, and then the extracellular fraction was obtained from the supernatant by filtration with a 0.20 μm membrane filter.…”

Section: Methodsmentioning

confidence: 99%

A GntR family transcription factor in Porphyromonas gingivalis regulates bacterial growth, acylpeptidyl oligopeptidase, and gingipains activity

Yang

Tan

Lei

et al. 2022

Molecular Oral Microbiology

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Porphyromonas gingivalis is a keystone pathogen for periodontitis. The function of the GntR family transcription factor is poorly studied in P. gingivalis. Numerous processes govern bacterial growth. The survival and pathogenicity of P. gingivalis depend heavily on its capacity to acquire amino acids as nutritional sources. In this investigation, a GntR transcription factor, pg1007, was identified in P. gingivalis, the deletion of which significantly inhibited bacterial growth. The mutant strain also exhibited an increased extracellular activity of gingipains and acylpeptidyl oligopeptidase (AOP). Global gene expression profiling revealed that the expression levels of 59 genes were significantly altered in the Δpg1007 mutant, with an upregulation in gene expression for AOP, ABC transporters, and some membrane proteins. In addition, His‐PG1007 protein was purified as a recombinant protein from Escherichia coli, and the conserved DNA sequence bound by it was determined using electrophoretic mobility shift assays and DNase I footprinting assays. Consequently, this study demonstrated that pg1007 is a crucial transcription factor in P. gingivalis and regulates the bacterial growth and activity of gingipains and AOP. These findings may enhance our understanding of the regulation of bacterial proliferation and protease activity in P. gingivalis.

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Section: Methodsmentioning

confidence: 99%

A GntR family transcription factor in Porphyromonas gingivalis regulates bacterial growth, acylpeptidyl oligopeptidase, and gingipains activity

Yang

Tan

Lei

et al. 2022

Molecular Oral Microbiology

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Illuminating the oral microbiome and its host interactions: tools and approaches for molecular microbiology studies

Merritt

Kreth

2022

FEMS Microbiology Reviews

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Advancements in DNA sequencing technologies within the last decade have stimulated an unprecedented interest in the human microbiome, largely due the broad diversity of human diseases found to correlate with microbiome dysbiosis. As a direct consequence of these studies, a vast number of understudied and uncharacterized microbes have been identified as potential drivers of mucosal health and disease. The looming challenge in the field is to transition these observations into defined molecular mechanistic studies of symbiosis and dysbiosis. In order to meet this challenge, many of these newly identified microbes will need to be adapted for use in experimental models. Consequently, this review presents a comprehensive overview of the molecular microbiology tools and techniques that have played crucial roles in genetic studies of the bacteria found within the human oral microbiota. Here, we will use specific examples from the oral microbiome literature to illustrate the biology supporting these techniques, why they are needed in the field, and how such technologies have been implemented. It is hoped that this information can serve as a useful reference guide to help catalyze molecular microbiology studies of the many new understudied and uncharacterized species identified at different mucosal sites in the body.

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Involvement of PG1037 in the repair of 8‐oxo‐7,8‐dihydroguanine caused by oxidative stress in Porphyromonas gingivalis

Dou,

Mishra,

Fletcher

2024

Molecular Oral Microbiology

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BackgroundThe PG1037 gene is part of the uvrA‐PG1037‐pcrA operon in Porphyromonas gingivalis. It encodes for a protein of unknown function upregulated under hydrogen peroxide (H2O2)‐induced oxidative stress. Bioinformatic analysis shows that PG1037 has a zinc‐finger motif, two peroxidase motifs, and one cytidylate kinase domain. The aim of this study is to characterize further the role of the PG1037 recombinant protein in the unique 8‐oxoG repair system in P. gingivalis.Materials and MethodsPG1037 recombinant proteins with deletions in the zinc‐finger or peroxidase motifs were created. Electrophoretic mobility shift assays were used to evaluate the ability of the recombinant proteins to bind 8‐oxoG‐containing oligonucleotides. Zinc binding, peroxidase, and Fenton reaction assays were used to assess the functional roles of the rPG1037 protein. A bacterial adenylate cyclase two‐bride assay was used to identify the partner protein of PG1037 in the repair of 8‐oxoG.ResultsThe recombinant PG1037 (rPG1037) protein carrying an N‐terminal His‐tag demonstrated an ability to recognize and bind 8‐oxoG‐containing oligonucleotide. In contrast to the wild‐type rPG1037 protein, the zinc‐finger motif deletion resulted in the loss of zinc and 8‐oxoG binding activities. A deletion of the peroxidase motif‐1 showed a decrease in peroxidase activity. Using a bacterial adenylate cyclase two‐hybrid system, there was no observed protein‐protein interaction of PG1037 with UvrA (PG1036), PcrA (PG1038), or mismatch repair system proteins.ConclusionsTaken together, the results show that PG1037 is an important member of a novel mechanism that recognizes and repairs oxidative stress‐induced DNA damage in P. gingivalis.

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PG1659 functions as anti‐sigma factor to extracytoplasmic function sigma factor RpoE in Porphyromonas gingivalis W83

Cited by 5 publications

References 63 publications

A GntR family transcription factor in Porphyromonas gingivalis regulates bacterial growth, acylpeptidyl oligopeptidase, and gingipains activity

A GntR family transcription factor in Porphyromonas gingivalis regulates bacterial growth, acylpeptidyl oligopeptidase, and gingipains activity

Illuminating the oral microbiome and its host interactions: tools and approaches for molecular microbiology studies

Involvement of PG1037 in the repair of 8‐oxo‐7,8‐dihydroguanine caused by oxidative stress in Porphyromonas gingivalis

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