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Related Experiment Videos

Microsatellite instability regulates transcription factor binding and gene expression.

Patricia Martin1, Katherine Makepeace, Stuart A Hill

  • 1Molecular Infectious Diseases Group, University of Oxford Department of Paediatrics, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom. martinp@necker.fr

Proceedings of the National Academy of Sciences of the United States of America
|February 25, 2005
PubMed
Summary

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Microsatellite instability in Neisseria meningitidis affects NadA expression by altering transcriptional regulation. Changes in TAAA repeats impact regulatory protein binding, influencing gene expression switching.

Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Microsatellites are repetitive DNA sequences prone to mutations.
  • In pathogenic bacteria, microsatellite instability drives phase variation, a mechanism for rapid phenotypic switching.
  • NadA, an adhesin from Neisseria meningitidis, exhibits phase-variable expression controlled by microsatellite repeats.

Purpose of the Study:

  • To investigate the impact of microsatellite repeat number variation on NadA gene transcription.
  • To explore the role of the Integration Host Factor (IHF) protein in regulating nadA expression.
  • To elucidate the regulatory mechanisms underlying NadA phase variation in Neisseria meningitidis.

Main Methods:

  • Analysis of TAAA repeat length variations in the nadA promoter region.

Related Experiment Videos

  • Electrophoretic mobility shift assays (EMSAs) to assess IHF binding to the nadA promoter.
  • Reporter gene assays to quantify nadA transcriptional activity.

    • Main Results:

    • Alterations in the number of TAAA repeats upstream of the nadA promoter were observed.
    • Changes in repeat length significantly affected the binding affinity of IHF to the nadA promoter.
    • Phase-variable transcription of nadA is directly influenced by IHF-promoter interactions modulated by repeat number.

    Conclusions:

    • Microsatellite instability in the nadA promoter region modulates IHF binding, thereby controlling NadA phase variation.
    • The regulation of nadA transcription involves a interplay between stochastic DNA mutations and protein-DNA interactions.
    • This study reveals a novel regulatory mechanism for bacterial gene expression, integrating mutational dynamics with classical transcriptional control.