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Parallel Measurement of Circadian Clock Gene Expression and Hormone Secretion in Human Primary Cell Cultures
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Enriching the Circadian Proteome.

Joseph S Takahashi1

  • 1Howard Hughes Medical Institute and Department of Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9111 USA.

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Circadian clocks control physiology and metabolism. New proteomics studies reveal extensive daily rhythms in the nuclear and phosphoproteome, expanding our understanding of these biological clocks.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chronobiology

Background:

  • Circadian clocks are endogenous biological timekeepers regulating daily physiological and metabolic processes.
  • Previous research identified circadian rhythms in the transcriptome, cistrome, and epigenome using genome-wide approaches.
  • The role of circadian regulation at the protein level, particularly the nuclear and phosphoproteome, remained less understood.

Purpose of the Study:

  • To investigate the extent of circadian regulation within the nuclear and phosphoproteome.
  • To identify specific proteins and phosphorylation sites exhibiting daily rhythms.
  • To provide a comprehensive proteomic view of circadian control in mammals.

Main Methods:

  • Utilized quantitative proteomics to analyze nuclear and phosphoproteomic samples.
  • Employed time-course experiments in mice to capture dynamic changes over a 24-hour cycle.
  • Applied sophisticated bioinformatics to identify rhythmically changing proteins and phosphosites.

Main Results:

  • Demonstrated widespread circadian rhythms in both the nuclear proteome and phosphoproteome.
  • Identified a significant number of proteins and phosphorylation events that oscillate with a circadian pattern.
  • Revealed extensive daily regulation at the protein and post-translational modification levels.

Conclusions:

  • Circadian clocks exert profound control over the nuclear and phosphoproteome.
  • Proteomics provides critical insights into the molecular mechanisms underlying circadian regulation.
  • These findings highlight the dynamic nature of the proteome in response to daily environmental cues.