Identification of critical amino acids in the DNA binding domain of LuxO: Lessons from a constitutive active LuxO

PLoS One. 2024 Sep 17;19(9):e0310444. doi: 10.1371/journal.pone.0310444. eCollection 2024.

Abstract

Quorum sensing plays a vital role in the environmental and host life cycles of Vibrio cholerae. The quorum-sensing circuit involves the consorted action of autoinducers, small RNAs, and regulatory proteins to control a plethora of physiological events in this bacterium. Among the regulatory proteins, LuxO is considered a low-cell-density master regulator. It is a homolog of NtrC, a two-component response regulator. NtrC belongs to an evolving protein family that works with the alternative sigma factor σ54 to trigger gene transcription. Structurally, these proteins comprise 3 domains: a receiver domain, a central AAA+ATPase domain, and a C-terminal DNA-binding domain (DBD). LuxO communicates with its cognate promoters by employing its DNA binding domain. In the present study, we desired to identify the critical residues in the DBD of LuxO. Our combined mutagenesis and biochemical assays resulted in the identification of eleven residues that contribute significantly to LuxO regulatory function.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / metabolism
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Domains
  • Quorum Sensing
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Vibrio cholerae* / genetics
  • Vibrio cholerae* / metabolism

Substances

  • Bacterial Proteins
  • Transcription Factors
  • DNA-Binding Proteins
  • Amino Acids
  • DNA, Bacterial

Grants and funding

This work was partly supported by grants from in-house OLP -193. SS, MK and HS received their fellowship from Council of Scientific and Industrial Research The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.