Deletion of the yqeK gene leads to the accumulation of Ap4A and reduced biofilm formation in Streptococcus mutans

Abstract

Diadenosine‐5′,5′′′‐P1, P4‐tetraphosphate (Ap4A) is a second messenger playing a crucial role in various life activities of bacteria. The increase of Ap4A expression is pleiotropic, resulting in an impairment in the formation of biofilm and other physiological functions in some bacteria. However, Ap4A function in Streptococcus mutans, an important pathogen related to dental caries, remains unknown. In this work, the Ap4A hydrolase, YqeK, was identified and characterized in S. mutans. Then, the effects of yqeK deletion on the growth, biofilm formation, and exopolysaccharide (EPS) quantification in S. mutans were determined by the assessment of the growth curve, crystal violet, and anthrone‐sulfuric acid, respectively, and visualized by microscopy. The results showed that the in‐frame deletion of the yqeK gene in S. mutans UA159 led to an increase in Ap4A levels, lag phase in the early growth, as well as decrease in biofilm formation and water‐insoluble exopolysaccharide production. Global gene expression profile showed that the expression of 88 genes was changed in the yqeK mutant, and among these, 42 were upregulated and 46 were downregulated when compared with the wild‐type S. mutans UA159. Upregulated genes were mainly involved in post‐translational modification, protein turnover, and chaperones, while downregulated genes were mainly involved in carbohydrate transport and metabolism. Important virulence genes related to biofilms, such as gtfB, gtfC, and gbpC, were also significantly downregulated. In conclusion, these results indicated that YqeK affected the formation of biofilms and the expression of biofilm‐related genes in S. mutans.

In Silico Molecular Docking of Phytochemicals of Murraya koenigii Against Streptococcus mutans

“…All four compounds show zero violations of the Lipinski rule. As only molecular docking is done in this study, further in vitro studies can be done on M. koenigii to find properties like antioxidant, antidiabetic, anticancer, and anti-inflammatory and can infuse the compounds of this plant into herbal medicines [14][15][16][17][18].…”

Regulatory mechanisms of exopolysaccharide synthesis and biofilm formation in Streptococcus mutans

“…Zheng et al . showed that the in-frame deletion of the yqeK gene in S. mutans causes an increase in the intracellular Ap4A levels, while biofilm formation and water-insoluble exopolysaccharides production are decreased [ 30 ]. The knockout of the yqeK gene leads to the downregulation of important virulence genes related to biofilms, such as gtfB , gtfC , and gtfD [ 30 ].…”

“…showed that the in-frame deletion of the yqeK gene in S. mutans causes an increase in the intracellular Ap4A levels, while biofilm formation and water-insoluble exopolysaccharides production are decreased [ 30 ]. The knockout of the yqeK gene leads to the downregulation of important virulence genes related to biofilms, such as gtfB , gtfC , and gtfD [ 30 ]. Although the mechanisms involved in the role of Ap4A need to be further investigated, these results suggest that it affects biofilm formation in S. mutans by affecting the expression of gtf s and the activity of Gtfs.…”

“… [ 25 ] cabPA Smu.1562 - [ 25 ] cabPB Smu.1708 - [ 25 ] yqeK Smu.1798c Deletion of yqeK inhibited exopolysaccharides synthesis and biofilm formation in S. mutans. [ 30 ] Regulation of S. mutans biofilm formation at transcriptional level epsR Smu.124 Deletion of epsR in S. mutans resulted in increased water-insoluble exopolysaccharides production, and upregulated GtfB protein content and activity. [ 31 ] rcrR Smu.921 Deletion of rcrR gene in S. mutans caused decreased biofilm formation ability.…”