The Oxidative Stress-Induced Hypothetical Protein PG-0686 in Porphyromonas gingivalis W83 Is a Novel Diguanylate Cyclase

Alexia D. Ximinies, Yuetan Dou, Arunima Mishra, Kangling Zhang, Champion Deivanayagam, Charles Wang, Hansel M. Fletcher

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


The survival/adaptation of Porphyromonas gingivalis to the inflammatory environment of the periodontal pocket requires an ability to overcome oxidative stress. Several functional classes of genes, depending on the severity and duration of the exposure, were induced in P. gingivalis under H2O2-induced oxidative stress. The PG0686 gene was highly upregulated under prolonged oxidative stress. PG0686, annotated as a hypothetical protein of unknown function, is a 60 kDa protein that carries several domains including hemerythrin, PAS10, and domain of unknown function (DUF)-1858. Although PG0686 showed some relatedness to several diguanylate cyclases (DGCs), it is missing the classical conserved, active site sequence motif (GGD[/E]EF), commonly observed in other bacteria. PG0686-related proteins are observed in other anaerobic bacterial species. The isogenic mutant P. gingivalis FLL361 (DPG0686::ermF) showed increased sensitivity to H2O2, and decreased gingipain activity compared to the parental strain. Transcriptome analysis of P. gingivalis FLL361 showed the dysregulation of several gene clusters/operons, known oxidative stress resistance genes, and transcriptional regulators, including PG2212, CdhR and PG1181 that were upregulated under normal anaerobic conditions. The intracellular level of c-di-GMP in P. gingivalis FLL361 was significantly decreased compared to the parental strain. The purified recombinant PG0686 (rPG0686) protein catalyzed the formation of c-di-GMP from GTP. Collectively, our data suggest a global regulatory property for PG0686 that may be part of an unconventional second messenger signaling system in P. gingivalis. Moreover, it may coordinately regulate a pathway(s) vital for protection against environmental stress, and is significant in the pathogenicity of P. gingivalis and other anaerobes. IMPORTANCE Porphyromonas gingivalis is an important etiological agent in periodontitis and other systemic diseases. There is still a gap in our understanding of the mechanisms that P. gingivalis uses to survive the inflammatory microenvironment of the periodontal pocket. The hypothetical PG0686 gene was highly upregulated under prolonged oxidative stress. Although the tertiary structure of PG0686 showed little relatedness to previously characterized diguanylate cyclases (DGCs), and does not contain the conserved GGD(/E)EF catalytic domain motif sequence, an ability to catalyze the formation of c-di-GMP from GTP is demonstrated. The second messenger pathway for c-di-GMP was previously predicted to be absent in P. gingivalis. PG0686 paralogs are identified in other anaerobic bacteria. Thus, PG0686 may represent a novel class of DGCs, which is yet to be characterized. In conclusion, we have shown, for the first time, evidence for the presence of c-di-GMP signaling with environmental stress protective function in P. gingivalis.

Original languageEnglish (US)
JournalMicrobiology Spectrum
Issue number2
StatePublished - Mar 2023


  • PG-0686
  • Porphyromonas gingivalis
  • c-di GMP
  • diguanylate cyclase
  • oxidative stress

ASJC Scopus subject areas

  • Physiology
  • Ecology
  • General Immunology and Microbiology
  • Genetics
  • Microbiology (medical)
  • Cell Biology
  • Infectious Diseases


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