title: Real-time analysis of the circadian oscillation of the Rev-Erb β promoter.
abstract: OBJECTIVE
  Rev-Erb β gene plays crucial roles in circadian rhythm, lipid and glucose metabolism, and several diseases. The molecular mechanisms of the transcriptional regulation of Rev-Erb β that generate and determine the phase of the circadian oscillation remain unclear.
  
  
  METHODS
  We analyzed the Rev-Erb β promoter by luciferase reporter assays, real-time bioluminescence monitoring assays and electrophoretic mobility shift assays.
  
  
  RESULTS
  Luciferase reporter assays indicated that only the 5' region and exon 1 have obvious promoter activity. Real-time bioluminescence monitoring assays revealed that E1, E2, E3, D boxes are important for maintenance of the amplitude of Rev-Erb β oscillation. Based on EMSA results, REV-ERBβ binds ROREs in the Bmal1 promoter region and inhibits Bmal1 promoter activity.
  
  
  CONCLUSIONS
  We provide direct evidence that three E-boxes and one D-box located in the first intron are crucial for the phase of circadian oscillation in Rev-Erb β expression and that the sequences upstream from its transcription start site function as a promoter with no circadian regulation. We also found that the E1 box affects the Rev-Erb β oscillation phase. Our results offer new insight into the role of Rev-Erb β in the circadian rhythm system.
authors.affiliation: Department of Neurology, Meitan General Hospital, Beijing, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: Ikuo
authors.initials: I
authors.lastname: Inoue
authors.forename: Megumi
authors.initials: M
authors.lastname: Kumagai
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Takahashi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Masaaki
authors.initials: M
authors.lastname: Ikeda
mesh_terms: D056930:ARNTL Transcription Factors / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D056926:CLOCK Proteins / Q000235:genetics; D002940:Circadian Rhythm; D017931:DNA Primers; D004742:Enhancer Elements, Genetic; D051379:Mice; D041681:NIH 3T3 Cells; D056953:Nuclear Receptor Subfamily 1, Group D, Member 1 / Q000235:genetics*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D056930:ARNTL Transcription Factors; C538961:Bmal1 protein, mouse; D017931:DNA Primers; D056953:Nuclear Receptor Subfamily 1, Group D, Member 1; D056926:CLOCK Proteins; C538944:Clock protein, mouse