5. RNA and DNA are shown in bold. GAR: 5-Phosphoribosyl glycinamide; FGAM: 5-phosphoribosyl-N-formylglycineamidine; FGAR: 1-(5′-Phosphoribosyl)-N-formylglycinamide; AICAR: 5′-phosphoribosyl-4-(N-succinocarboxamide)-5-aminoimidazole; AIR: 1-(5′-Phophoribosyl)-5-aminoimidazole; CAIR: 5′P-Ribosyl-4-carboxy-5-aminoimidazole; SAICAR: 5′P-Ribosyl-4-(N-succinocarboximide)-5-aminoimidazole; selleck screening library FAICAR: 1 (5′-Phosphoribosyl)-5-formamido-4-imidazole carboxamide. Stress proteins The ability of the community to provide physiologic support to constituent species might result in P. gingivalis experiencing lower levels of environmental stress than occurs in monoculture. Consistent with
this concept, community derived P. gingivalis showed a significant reduction in abundance of DNA repair proteins (PGN0333, RadA; PGN0342, Ung; PGN0367, Xth; PGN1168, MutS; PGN1316, UvrA; PGN1388, LigA; PGN1567, RecF; PGN1585, UvrB; PGN1712, Nth; PGN1714, Mfd; PGN1771, Pol1). DNA repair genes are generally induced in the presence of damaged DNA see more [41], and lower abundance of DNA repair proteins is consistent with the monoculture experiencing more DNA damage than P. gingivalis in the three species community where the presence of the partner organisms provides protection against DNA damage. Only two stress proteins showed increased abundance, and then
only 30% increases, the molecular chaperone DnaK (PGN1208) and a PhoH family protein possibly involved in oxidation protection (PGN0090). Role of the differentially regulated P. gingivalis protein HmuR To begin to test the functional relevance of proteins identified as differentially regulated in the three species community, we undertook a mutational analysis. For this purpose it was important to target Bumetanide a protein that directly effectuates a biological LY411575 function and lacks homologs in the genome. HmuR, a major hemin uptake
protein, and potential adhesin [42], was selected. As shown in Fig. 7A, while wild type P. gingivalis cells are abundant within a S. gordonii-F. nucleatum-P. gingivalis community, P. gingivalis cells lacking HmuR are deficient in community formation. Biovolume analysis showed a 70% reduction in community formation by the HmuR mutant (Fig. 7C). Furthermore, this effect was specific for the three species community as a decrease in accumulation by the HmuR deficient mutant was not observed in monospecies biofilms, or in two species communities of P. gingivalis with either S. gordonii or F. nucleatum (Fig. 7B, D–G). Hence loss of HmuR, that is up-regulated by P. gingivalis when the organism is associated with S. gordonii and F. nucleatum, results in a phenotype that is restricted to three species community formation. P. gingivalis cells were first cultured in hemin excess, under which conditions the hmu operon is expressed at a basal level [42]. As the three species model system involves metabolically quiescent P.