CBP/p300 is a cell type-specific modulator of CLOCK/BMAL1-mediated transcription.

Background: Previous studies have demonstrated tissue-specific regulation of the rhythm of circadian transcription, suggesting that transcription factor complex CLOCK/BMAL1, essential for maintaining circadian rhythm, regulates transcription in a tissue-specific manner. To further elucidate the mechanism of the cell type-specific regulation of transcription by CLOCK/BMAL1 at the molecular level, we investigated roles of CBP/p300 and tissue-specific cofactors in CLOCK/BMAL1-mediated transcription.

Results: As shown previously, CBP/p300 stimulates CLOCK/BMAL1-mediated transcription in COS-1 cells.

Characterization of the promoter of the mouse preproorexin gene.

The neuropeptide Orexin is involved in the regulation of the sleep-awake cycle and feeding behavior. We isolated a 22-kb genomic clone containing the 5' flanking region of the mouse Orexin promoter. We determined that the transcription start site (+1) is located 96 nucleotides upstream of the initiation codon.

A BMAL1 mutant with arginine 91 substituted with alanine acts as a dominant negative inhibitor.

Basic-Helix-Loop-Helix-Per-Arnt-Sim (bHLH-PAS) transcription factor, Brain-Muscle-Arnt-Like-protein 1 (BMAL1), forms a heterodimer with the CLOCK protein. The BMAL1/CLOCK complex binds to a specific DNA sequence and plays an essential role in the generation of the circadian rhythm. The basic region of BMAL1 contains an E-R-X-R motif that is highly conserved among basic-helix-loop-helix (bHLH) transcription factors that bind to the E-box transcription element, and is thus thought to constitute a structure required for recognition of this DNA sequence.