title: Survival of an Aggregatibacter actinomycetemcomitans quorum sensing luxS mutant in the mouths of Rhesus monkeys: insights into ecological adaptation.
abstract: Experiments were designed to explore a prominent autoinducer-2 (AI-2) producing gene (luxS) related to colonization and survival of Aggregatibacter actinomycetemcomitans, a low abundance member of the indigenous flora, that forms a key component of the dysbiotic flora in localized aggressive periodontitis. The luxS gene was disrupted in a primate strain of A. actinomycetemcomitans before implantation into the oral cavity of Rhesus monkeys (Rh). The colonization efficiency of the luxS mutant (RhAa-VS4) was compared with the parental wild-type strain (RhAa3) (positive control) and a ltxA mutant (RhAa-VS2) (negative control). The in vivo results showed that the luxS mutation had minimal impact on A. actinomycetemcomitans colonization compared with the wild-type RhAa3 strain. In vitro studies revealed that there was a significant upregulation of attachment-related genes aae, apiA, and flp in the RhAa-VS4 strain compared with RhAa3. Biofilm forming ability was also significantly increased in the RhAa-VS4 strain compared with RhAa3, whereas the AI-2 signal was ablated. The exogenous addition of the AI-2 precursor dihydroxy pentanedione allowed the RhAa-VS4 strain to achieve RhAa3 biofilm levels. This is the first primate study to test the relevance of LuxS in vivo. In vitro assessment suggests that in vivo survival of the RhAa-VS4 strain was due to the production of signaling AI-2 molecules derived from other members of the flora as well as the upregulation of genes related to attachment and biofilm formation.
authors.affiliation: Department of Oral Biology, Rutgers School of Dental Medicine, Rutgers University, Newark, NJ, USA.
authors.forename: Senthil K
authors.initials: SK
authors.lastname: Velusamy
authors.affiliation: Department of Oral Biology, Rutgers School of Dental Medicine, Rutgers University, Newark, NJ, USA.
authors.forename: Vandana
authors.initials: V
authors.lastname: Sampathkumar
authors.affiliation: Department of Oral Biology, Rutgers School of Dental Medicine, Rutgers University, Newark, NJ, USA.
authors.forename: Dipti
authors.initials: D
authors.lastname: Godboley
authors.affiliation: Department of Oral Biology, Rutgers School of Dental Medicine, Rutgers University, Newark, NJ, USA.
authors.forename: Daniel H
authors.identifier: 0000-0001-6242-6451
authors.initials: DH
authors.lastname: Fine
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics*; D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics* / Q000502:physiology*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D018441:Biofilms; D013437:Carbon-Sulfur Lyases / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006714:Homoserine / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism; D007783:Lactones / Q000378:metabolism; D008253:Macaca mulatta; D050296:Microbial Viability; D009055:Mouth / Q000382:microbiology*; D009154:Mutation; D053038:Quorum Sensing / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D007783:Lactones; C091088:N-octanoylhomoserine lactone; D006714:Homoserine; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria
keywords: Rhesus monkey; autoinducer-2; biofilm; oral colonization; quorum sensing