Transcriptional repressor PRR5 directly regulates clock-output pathways

Proc Natl Acad Sci U S A. 2012 Oct 16;109(42):17123-8. doi: 10.1073/pnas.1205156109. Epub 2012 Oct 1.

Abstract

The circadian clock is an endogenous time-keeping mechanism that enables organisms to adapt to external daily cycles. The clock coordinates biological activities with these cycles, mainly through genome-wide gene expression. However, the exact mechanism underlying regulation of circadian gene expression is poorly understood. Here we demonstrated that an Arabidopsis PSEUDO-RESPONSE REGULATOR 5 (PRR5), which acts in the clock genetic circuit, directly regulates expression timing of key transcription factors involved in clock-output pathways. A transient expression assay and ChIP-quantitative PCR assay using mutated PRR5 indicated that PRR5 associates with target DNA through binding at the CCT motif in vivo. ChIP followed by deep sequencing coupled with genome-wide expression profiling revealed the direct-target genes of PRR5. PRR5 direct-targets include genes encoding transcription factors involved in flowering-time regulation, hypocotyl elongation, and cold-stress responses. PRR5-target gene expression followed a circadian rhythm pattern with low, basal expression from noon until midnight, when PRR9, PRR7, and PRR5 were expressed. ChIP-quantitative PCR assays indicated that PRR7 and PRR9 bind to the direct-targets of PRR5. Genome-wide expression profiling using a prr9 prr7 prr5 triple mutant suggests that PRR5, PRR7, and PRR9 repress these targets. Taken together, our results illustrate a genetic network in which PRR5, PRR7, and PRR9 directly regulate expression timing of key transcription factors to coordinate physiological processes with daily cycles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / physiology*
  • Chromatin Immunoprecipitation
  • Circadian Clocks / physiology
  • Circadian Rhythm / physiology*
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant / physiology*
  • High-Throughput Nucleotide Sequencing
  • Mutation / genetics
  • Polymerase Chain Reaction
  • Repressor Proteins / physiology*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcription Factors / physiology

Substances

  • Arabidopsis Proteins
  • PRR5 protein, Arabidopsis
  • PRR7 protein, Arabidopsis
  • PRR9 protein, Arabidopsis
  • Repressor Proteins
  • Transcription Factors

Associated data

  • GEO/GSE36360
  • GEO/GSE36361