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Related Experiment Video

Updated: Sep 2, 2025

Purification of a High Molecular Mass Protein in Streptococcus mutans
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AhrC Negatively Regulates Streptococcus mutans Arginine Biosynthesis.

Meiling Jing1,2, Ting Zheng1, Tao Gong1

  • 1State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan Universitygrid.13291.38, Chengdu, China.

Microbiology Spectrum
|August 8, 2022
PubMed
Summary

The transcription factor AhrC negatively regulates arginine biosynthesis and biofilm formation in Streptococcus mutans. This discovery clarifies arginine metabolism regulation in this cariogenic bacterium.

Keywords:
Streptococcus mutansbiofilm(s)gene expressionmicrobial geneticstranscription factor(s)

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Streptococcus mutans is a key pathogen in human dental caries.
  • Arginine metabolism is vital for bacterial growth, but its regulation in S. mutans is not fully understood.
  • The role of transcription factors in controlling arginine metabolism in S. mutans requires investigation.

Purpose of the Study:

  • To investigate the function and regulatory mechanism of ArgR family transcription factors in S. mutans.
  • To identify a specific transcription factor, AhrC, and elucidate its role in arginine metabolism and biofilm formation.

Main Methods:

  • Construction and analysis of ahrC in-frame deletion and overexpression strains.
  • Measurement of intracellular arginine content, biofilm biomass, and exopolysaccharide (EPS) production.
  • Global gene expression profiling (RNA sequencing) and analysis of differentially expressed genes.
  • Electrophoretic mobility shift assay (EMSA) and DNase I footprinting to identify DNA binding sites.
  • β-galactosidase activity assays to confirm regulatory function.

Main Results:

  • The ahrC deletion strain showed increased intracellular arginine and slow growth, while overexpression reduced arginine content.
  • Overexpression of ahrC led to decreased biofilm biomass, reduced water-insoluble EPS, and altered biofilm structures.
  • Gene expression analysis revealed significant upregulation of arginine biosynthesis genes (e.g., argJ, argB, argC) in the deletion strain and downregulation in the overexpression strain.
  • AhrC was confirmed to bind specific DNA sequences and act as a negative regulator of gene expression.

Conclusions:

  • AhrC is a critical transcription factor that negatively regulates arginine biosynthesis and biofilm formation in S. mutans.
  • AhrC directly controls the expression of arginine biosynthesis genes, including N-acetyl-gamma-glutamyl-phosphate reductase (argC).
  • These findings enhance the understanding of arginine metabolism regulation and its impact on biofilm development in Streptococcus mutans.