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The Phospho-Code Determining Circadian Feedback Loop Closure and Output in Neurospora.

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Summary

This study reveals a specific phosphorylation code on White Collar-1 and White Collar-2 proteins that regulates circadian rhythm feedback loops in fungi and animals, providing mechanistic insights into circadian output.

Keywords:
C-boxDNA bindingFRQWC-1WC-2ccgsclock-controlled genesfeedback loopfrq transcriptionphosphorylation

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Area of Science:

  • Chronobiology
  • Molecular Biology
  • Biochemistry

Background:

  • Circadian rhythms rely on negative feedback loops involving transcriptional repressors.
  • The phosphorylation of transcriptional activators like the White Collar complex (WCC) is proposed to mediate this repression, but mechanisms are unclear.

Purpose of the Study:

  • To elucidate the phosphorylation-dependent mechanisms governing circadian negative feedback and output.
  • To identify the specific phosphosites and phosphorylation patterns regulating the White Collar complex (WCC).

Main Methods:

  • Phosphoproteomic analysis to map phosphosites on WC-1 and WC-2 proteins.
  • Investigating the role of identified phosphosites in WCC activity and gene regulation.

Main Results:

  • Identified 80 phosphosites on WC-1 and 15 on WC-2.
  • Elucidated a time-of-day-specific phosphorylation code on WC-1 and WC-2 essential for circadian repression.
  • Demonstrated that combinatorial phosphorylation controls rhythmic WCC binding to target gene promoters.

Conclusions:

  • Established a mechanistic understanding of how WCC phosphorylation drives circadian negative feedback.
  • Provided insights into the regulation of circadian output gene expression through phosphorylation-mediated WCC binding.