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Related Experiment Videos

Second messenger and Ras/MAPK signalling pathways regulate CLOCK/CYCLE-dependent transcription.

Frank Weber1, Hsiu-Cheng Hung, Christian Maurer

  • 1Biochemie-Zentrum Heidelberg, Ruprecht-Karls Universität Heidelberg, Heidelberg, Germany.

Journal of Neurochemistry
|June 30, 2006
PubMed
Summary
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This study identifies key kinases regulating the CLOCK/CYCLE complex, essential for circadian rhythms. Protein kinase A, CaMK II, and MAPK directly phosphorylate CLOCK, controlling circadian gene expression.

Area of Science:

  • Molecular Biology
  • Chronobiology
  • Biochemistry

Background:

  • The CLOCK/CYCLE complex is central to circadian clock function in Drosophila and mammals.
  • Protein phosphorylation is crucial for regulating the molecular oscillator.
  • Kinases and signaling pathways controlling CLOCK/CYCLE activity are largely unknown.

Purpose of the Study:

  • To identify kinases and signaling pathways that regulate CLOCK/CYCLE function.
  • To elucidate the mechanisms by which these kinases impact circadian gene expression.

Main Methods:

  • Chemical screen of kinase inhibitors using a cell culture reporter assay.
  • Analysis of constitutively active kinases.
  • In vitro phosphorylation assays.

Related Experiment Videos

Main Results:

  • Cyclic nucleotide/protein kinase A (PKA), calcium/calmodulin-dependent kinase (CaMK) II, and Ras/mitogen-activated protein kinase (MAPK) were identified as regulators of CLOCK/CYCLE activity.
  • CaMK II and p42 MAPK (ERK2) directly phosphorylate CLOCK in vitro.
  • These kinases regulate CLOCK/CYCLE-dependent transcription through direct phosphorylation of CLOCK.

Conclusions:

  • PKA, CaMK II, and MAPK are key regulators of the Drosophila and mammalian circadian clock.
  • Direct phosphorylation of CLOCK by CaMK II and MAPK is a critical mechanism for controlling circadian transcription.