cyclic-gmp and Periodontal-Diseases

Periodontal disease (PD) is a progressive inflammatory condition characterized by degradation of the gingival epithelium, periodontal ligament, and alveolar bone ultimately resulting in tooth loss. Treponema denticola is a keystone periopathogen that contributes to immune dysregulation and direct tissue destruction. As periodontal disease develops, T. denticola must adapt to environmental, immunological and physiochemical changes in the subgingival crevice. Treponema denticola produces bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP), an important regulatory nucleotide. While T. denticola encodes several putative diguanylate cyclases (DGCs), none have been studied and hence the biological role of c-di-GMP in oral treponemes remains largely unexplored. Here, we demonstrate that the T. denticola open reading frame, TDE0125, encodes a functional DGC designated as DgcA (Diguanylate cyclase A). The dgcA gene is universal among T. denticola isolates, highly conserved and is a stand-alone GGEEF protein with a GAF domain. Recombinant DgcA converts GTP to c-di-GMP using either manganese or magnesium under aerobic and anaerobic reaction conditions. Size exclusion chromatography revealed that DgcA exists as a homodimer and in larger oligomers. Site-directed mutagenesis of residues that define the putative inhibitory site of DgcA suggest that c-di-GMP production is allosterically regulated. This report is the first to characterize a DGC of an oral treponeme.

Topics: Bacterial Proteins; Cyclic GMP; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Guanosine Triphosphate; Humans; Mutagenesis, Site-Directed; Periodontal Diseases; Phosphorus-Oxygen Lyases; Phylogeny; Protein Domains; Recombinant Proteins; Sequence Analysis, DNA; Treponema denticola

Topics: Animals; Biology; Cell Transformation, Viral; Collagen; Cyclic AMP; Cyclic GMP; Fibroblasts; Glucocorticoids; Humans; Mice; Periodontal Diseases; Periodontium; Rats; Receptors, Cell Surface; Steroids