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Watt, R. M.

Publications and source records attributed to Watt, R. M..

2 recordsLinked to original sources

Diversity of Treponema denticola and other oral treponeme lineages in subjects with periodontitis and gingivitis

More than 75 species/species-level phylotypes of taxa belonging to the genus Treponema inhabit the human oral cavity. Treponema denticola is commonly associated with periodontal disease, but the etiological roles and ecological distributions of other oral treponemes remain more obscure. Here, we compared the clinical distributions of phylogroup 1 and 2 oral treponemes in subgingival plaque sampled from Chinese subjects with periodontitis (n=10) and gingivitis (n=8), via the sequence analysis of the highly-conserved pyrH housekeeping gene. Two PCR primer sets that respectively targeted oral phylogroup 1 and 2 treponeme pyrH genes were used to construct plasmid clone-amplicon libraries for each subject, which were sequenced for bioinformatic analysis. 1,204 quality-filtered, full-length pyrH gene sequences were obtained from the cohort (median: 61.5; range: 59-83 of cloned pyrH sequences per subject), which were assigned to 34 pyrH genotypes (pyrH001-pyrH034; 97% sequence-identity cut-off). 18 pyrH genotypes (536 pyrH sequences) corresponded to phylogroup 1 treponeme taxa (including Treponema vincentii, Treponema medium). 16 pyrH genotypes (668 pyrH sequences) corresponded to T. denticola and other phylogroup 2 treponemes. Periodontitis subjects contained a greater diversity of phylogroup 2 pyrH genotypes, compared to gingivitis subjects (Mann-Whitney U-test). One T. denticola pyrH genotype (pyrH001) was highly prevalent: detected in 10/10 periodontitis and 6/8 gingivitis subjects. Several subjects harboured multiple T. denticola pyrH genotypes. Non-metric multidimensional scaling and PERMANOVA tests revealed no significant differences in overall pyrH genotype compositions between periodontitis and gingivitis subjects. In conclusion, our results clearly show that subjects with periodontitis and gingivitis commonly harbour highly-diverse oral treponeme communities.

microbiology↗

The Ps and Qs of alarmone synthesis in Staphylococcus aureus

During the stringent response, bacteria synthesize guanosine-3,5-bis(diphosphate) (ppGpp) and guanosine-5-triphosphate 3-diphosphate (pppGpp), which act as secondary messengers to promote cellular survival and adaptation. (p)ppGpp alarmones are synthesized and/or hydrolyzed by proteins belonging to the RelA/SpoT Homologue (RSH) family. Many bacteria also encode small alarmone synthetase (SAS) proteins (e.g. RelP, RelQ) which may also be capable of synthesizing a third alarmone: guanosine-5-phosphate 3-diphosphate (pGpp). Here, we report the biochemical properties of the Rel (RSH), RelP and RelQ proteins from Staphylococcus aureus (Sa-Rel, Sa-RelP, Sa-RelQ, respectively). Sa-Rel synthesized pppGpp more efficiently than ppGpp, but lacked the ability to produce pGpp. However, Sa-Rel efficiently hydrolyzed all three alarmones in a Mn(II) ion-dependent manner. The removal of the C-terminal regulatory domain of Sa-Rel increased its rate of (p)ppGpp synthesis ca. 10-fold, but had negligible effects on its rate of (pp)pGpp hydrolysis. Sa-RelP and Sa-RelQ efficiently synthesized pGpp in addition to pppGpp and ppGpp. The alarmone-synthesizing abilities of Sa-RelQ, but not Sa-RelP, were allosterically-stimulated by the addition of pppGpp, ppGpp or pGpp. The respective (pp)pGpp-synthesizing activities of Sa-RelP/Sa-RelQ were compared and contrasted with SAS homologues from Enterococcus faecalis (Ef-RelQ) and Streptococcus mutans (Sm-RelQ, Sm-RelP). Results indicated that EF-RelQ, Sm-RelQ and Sa-RelQ were functionally-equivalent; but exhibited considerable variations in their respective biochemical properties, and the degrees to which alarmones and single-stranded RNA molecules allosterically stimulated their respective alarmone-synthesizing activities. The respective (pp)pGpp-synthesizing capabilities of Sa-RelP and Sm-RelP proteins were inhibited by pGpp, ppGpp and pppGpp. Our results support the premise that RelP and RelQ proteins may synthesize pGpp in addition to (p)ppGpp within S. aureus and other Gram-positive bacterial species.

biochemistry↗