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Related Experiment Videos

DNA target sequence and FNR-dependent gene expression.

Colin Scott1, Jonathan D Partridge, James R Stephenson

  • 1Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, UK.

FEBS Letters
|April 23, 2003
PubMed
Summary
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FNR proteins regulate gene transcription in response to oxygen levels. The non-conserved tetrad (NCT) sequence within their DNA targets significantly impacts the dynamic range of this oxygen-sensing switch.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • FNR (fumarate and nitrate reduction) proteins are crucial global transcription regulators in bacteria.
  • They control gene expression in response to environmental oxygen availability.
  • FNR proteins bind to a specific DNA target sequence: TTGATN(1)N(2)N(3)N(4)ATCAA, where N(1-4) is a non-conserved tetrad (NCT).

Purpose of the Study:

  • To investigate the role of the NCT sequence in FNR-dependent gene regulation.
  • To determine how variations in the NCT sequence affect FNR's ability to control transcription.
  • To understand the impact of NCT on the dynamic range of the FNR regulatory switch.

Main Methods:

  • Bioinformatic analysis of 68 known and predicted FNR binding sites in the Escherichia coli K12 genome.

Related Experiment Videos

  • In vivo assessment of FNR-dependent transcription using class II and class I model promoters with engineered NCT sequences.

    • Main Results:

    • Analysis revealed a bias towards Adenine (A) or Thymine (T) at positions N(2) and N(3) of the NCT sequence in E. coli.
    • Altering the NCT sequence did not affect basal promoter activity.
    • Changes in the NCT sequence significantly altered anaerobic induction of transcription, by up to tenfold.

    Conclusions:

    • The NCT sequence is not essential for basal FNR activity but is a critical determinant of transcriptional regulation.
    • The NCT sequence plays a fundamental role in establishing the dynamic range or induction magnitude of the FNR regulatory switch.
    • Understanding NCT sequence variations provides insights into fine-tuning bacterial responses to oxygen.