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A cryptic transcription factor regulates Caulobacter adhesin development.

Maeve McLaughlin1, David M Hershey2, Leila M Reyes Ruiz3

  • 1Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States of America.

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Summary

Researchers identified RtrC, a novel regulator controlling bacterial adhesion in Caulobacter crescentus. RtrC directly represses hfiA, activating holdfast development and influencing surface attachment and biofilm formation.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Adhesion

Background:

  • Alphaproteobacteria utilize a polar adhesin, the holdfast, for surface attachment and biofilm formation.
  • Holdfast biogenesis in Caulobacter crescentus is indirectly regulated by two-component signal transduction (TCS) proteins, controlling expression of the holdfast inhibitor HfiA.

Purpose of the Study:

  • To identify direct regulators of hfiA downstream of the adhesion TCS system.
  • To elucidate the molecular mechanism and functional role of novel regulators in bacterial adhesion.

Main Methods:

  • Genetic selection to identify direct hfiA regulators.
  • Chromatin immunoprecipitation sequencing (ChIP-seq) to identify RtrC binding sites.
  • Assays for surface adherence (cheesecloth binding) and pellicle biofilm formation.

Main Results:

  • The hypothetical gene rtrC was identified as a direct regulator of hfiA.
  • RtrC binds to numerous chromosomal sites, including the hfiA promoter, repressing its transcription.
  • RtrC and SpdR form an OR-gated type I coherent feedforward loop regulating hfiA.
  • Mutations in rtrC diminished surface adherence and pellicle biofilm formation.

Conclusions:

  • RtrC is a novel, DNA-binding transcriptional repressor that directly activates holdfast development.
  • The identified regulatory network, involving RtrC and SpdR, provides a mechanism for buffering gene expression against signal fluctuations.
  • RtrC plays a crucial role in Caulobacter crescentus surface colonization and biofilm development.