Journal Article

Acetylation of glucosyltransferases regulates Streptococcus mutans biofilm formation and virulence

Li, Yuqing, Pan, Yangyang, Gong, Tao, Ma, Qizhao, Chen, Yang, Zou, Jing, Yu, Shuxing, Liu, Yaqi, Huang, Jun

2021

Overview

Lysine acetylation is a frequently occurring post-translational modification (PTM), emerging as an important metabolic regulatory mechanism in prokaryotes. This process is achieved enzymatically by the protein acetyltransferase (KAT) to specifically transfer the acetyl group, or non-enzymatically by direct intermediates (acetyl phosphate or acetyl-CoA). Although lysine acetylation modification of glucosyltransferases (Gtfs), the important virulence factor in Streptococcus mutans , was reported in our previous study, the KAT has not been identified. Here, we believe that the KAT ActG can acetylate Gtfs in the enzymatic mechanism. By overexpressing 15 KATs in S . mutans , the synthesized water-insoluble extracellular polysaccharides (EPS) and biofilm biomass were measured, and KAT ( actG ) was identified. The in-frame deletion mutant of actG was constructed to validate the function of actG . The results showed that actG could negatively regulate the water-insoluble EPS synthesis and biofilm formation. We used mass spectrometry (MS) to identify GtfB and GtfC as the possible substrates of ActG. This was also demonstrated by in vitro acetylation assays, indicating that ActG could increase the acetylation levels of GtfB and GtfC enzymatically and decrease their activities. We further found that the expression level of actG in part explained the virulence differences in clinically isolated strains. Moreover, overexpression of actG in S . mutans attenuated its cariogenicity in the rat caries model. Taken together, our study demonstrated that the KAT ActG could induce the acetylation of GtfB and GtfC enzymatically in S . mutans , providing insights into the function of lysine acetylation in bacterial virulence and pathogenicity.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Acetyl phosphate / Acetylation / Acetyltransferase / Acetyltransferases - metabolism / Animals / Bacteria / Biofilms / Biology and Life Sciences / Biomass / Cariogenicity / Causes of / Deletion mutant / Dental caries / Development and progression / Female / Gene deletion / Genetic aspects / Glucosyltransferases - metabolism / Glycosyltransferases / Health aspects / Intermediates / Lysine / Lysine - metabolism / Mass spectrometry / Mass spectroscopy / Medicine and Health Sciences / Microbial mats / Pathogenicity / Pathogens / Physical Sciences / Physiological aspects / Polysaccharides / Post-translation / Prokaryotes / Proteins / Rats / Rats, Sprague-Dawley / Regulatory mechanisms (biology) / Research and Analysis Methods / Saccharides / Scanning electron microscopy / Streptococcus / Streptococcus infections / Streptococcus mutans / Streptococcus mutans - pathogenicity / Streptococcus mutans - physiology / Substrates / Sucrose / Virulence / Virulence - physiology / Virulence factors