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Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
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Rhodobacter sphaeroides CarD Negatively Regulates Its Own Promoter.

Kemardo K Henry1, Wilma Ross1, Richard L Gourse1

  • 1Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, USA.

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|June 21, 2021
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Summary

The transcription factor CarD activates many promoters in Rhodobacter sphaeroides but inhibits its own promoter. This autoregulation involves CarD binding to RNA polymerase and DNA, reducing full-length mRNA synthesis by increasing abortive transcripts.

Keywords:
CarDR. sphaeroides promotersnegative autoregulationpromoter escapetranscription factortranscription inhibition

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Many bacterial promoters, particularly in Rhodobacter sphaeroides, lack a conserved thymine at position -7 of the -10 element.
  • The transcription factor CarD was previously shown to activate promoters lacking this -7T base, compensating for low basal activity.
  • The promoter for carD itself in R. sphaeroides was uncharacterized regarding its regulation by CarD.

Purpose of the Study:

  • To investigate the regulation of the carD promoter by CarD in Rhodobacter sphaeroides.
  • To elucidate the mechanism by which CarD affects its own promoter activity.
  • To understand how CarD's interaction with RNA polymerase and DNA influences transcription initiation and elongation.

Main Methods:

  • Characterization of the carD promoter's basal activity in R. sphaeroides.
  • In vitro transcription assays to assess the effect of CarD on carD promoter activity.
  • Analysis of abortive transcript formation and promoter escape in the presence of CarD.

Main Results:

  • The carD promoter exhibits high basal activity despite lacking -7T, dependent on an extended -10 element (TGn) and specific spacer features.
  • CarD inhibits, rather than activates, its own promoter activity.
  • CarD negatively autoregulates by increasing abortive transcription, limiting promoter escape and full-length mRNA synthesis.

Conclusions:

  • CarD employs a unique negative autoregulation mechanism involving increased abortive initiation at its own promoter.
  • This mechanism differs from classical repression and other known allosteric inhibition modes.
  • CarD's effect on transcription (activation or inhibition) is promoter-specific, depending on DNA sequence and kinetic properties.