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Related Experiment Videos

The E47 transcription factor negatively regulates CD5 expression during thymocyte development.

Yang Yang1, Christopher H Contag, Dean Felsher

  • 1Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.

Proceedings of the National Academy of Sciences of the United States of America
|March 6, 2004
PubMed
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Transcription factor E47 negatively regulates CD5 expression in developing T cells. Decreased E47 levels during thymocyte maturation are critical for increased CD5 expression, revealing a novel regulatory mechanism.

Area of Science:

  • Immunology
  • Molecular Biology
  • Developmental Biology

Background:

  • CD5 expression increases during thymocyte maturation.
  • CD5 expression is regulated by a specific promoter with transcription factor binding sites.
  • The role of E47 transcription factor in CD5 regulation was previously unknown.

Purpose of the Study:

  • To investigate the role of the E47 transcription factor and the kappa E2 site in CD5 regulation during thymocyte development.
  • To elucidate the molecular mechanisms controlling progressive CD5 expression in maturing thymocytes.

Main Methods:

  • Site-directed mutagenesis of the CD5 promoter's kappa E2 site.
  • Electrophoretic mobility shift assays (EMSAs) to assess E47 binding.
  • Overexpression studies of E47 in T cell lines.

Related Experiment Videos

  • High-dimensional fluorescence-activated cell sorting (FACS) analysis of primary thymocytes.
  • In vivo and in vitro stimulation of thymocytes.

    • Main Results:

    • Mutation of the kappa E2 site increased CD5 promoter activity.
    • E47 transcription factor binds to the kappa E2 site.
    • E47 overexpression inhibited CD5 expression.
    • Intracellular E47 levels decreased as surface CD5 expression increased during thymocyte development.
    • E47 was down-regulated and CD5 up-regulated following pre-T cell receptor stimulation in vivo and in vitro.

    Conclusions:

    • E47 transcription factor negatively regulates CD5 expression via the kappa E2 site in the CD5 promoter.
    • Decreased E47 levels in response to developmental signals are essential for the progressive increase in CD5 expression during thymocyte maturation.