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E-box function in a period gene repressed by light.

Daniela Vallone1, Srinivas Babu Gondi, David Whitmore

  • 1Max-Planck-Institut für Entwicklungsbiologie, Spemannstrasse 35-39, D-72076 Tübingen, Germany.

Proceedings of the National Academy of Sciences of the United States of America
|March 17, 2004
PubMed
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We discovered a zebrafish period (per) gene, zfper4, that is repressed by light, unlike most per genes. This finding reveals flexibility in how E-box elements control circadian gene expression and light responsiveness.

Area of Science:

  • Chronobiology
  • Molecular Biology
  • Genetics

Background:

  • Light synchronizes circadian clocks in most organisms.
  • Light pulses phase-shift circadian clocks and induce period (per) gene expression in vertebrates.

Purpose of the Study:

  • To clone and characterize a novel zebrafish per gene, zfper4.
  • To investigate the role of E-box elements in zfper4 regulation and light responsiveness.
  • To develop a reporter assay for studying light-entrainable clocks.

Main Methods:

  • Cloning of the zebrafish zfper4 gene.
  • Development of an in vivo luciferase reporter assay in cells with a light-entrainable clock.
  • High-definition bioluminescence to measure clock phase-shifting.
  • Site-directed mutagenesis of E-box elements in the zfper4 promoter.

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Main Results:

  • Identified zfper4, a zebrafish per gene repressed by light.
  • Demonstrated that four E-box elements in the zfper4 promoter are critical for basal expression, rhythmicity, and light repression.
  • Showed that these E-box elements, in a heterologous promoter, drive phase-advanced rhythms lacking light repression.

Conclusions:

  • The zfper4 gene exhibits unique light-repression properties.
  • E-box elements confer flexibility in circadian rhythm phase and light responsiveness depending on promoter context.
  • The developed reporter assay is valuable for studying light-mediated circadian clock regulation.