Regulation of the PAI-1 promoter by circadian clock components: differential activation by BMAL1 and BMAL2.
Circadian variation in plasminogen activator inhibitor-1 (PAI-1) production likely contributes to increased risk of myocardial infarction and decreased efficacy of thrombolytic therapy during the morning. In this study, we characterize the abilities of fundamental molecular components of intrinsic circadian clocks to regulate the human PAI-1 promoter in transfected endothelial cells. Both CLOCK:BMAL1 and CLOCK:BMAL2 heterodimers activate the PAI-1 promoter through requisite proximal (-565 to -560 bp) and distal (-680 to -675 bp) E-box enhancers. Although the distal E-box overlaps the 4G/5G polymorphism of the PAI-1 promoter, allelic variation at this site does not influence CLOCK:BMAL1-and CLOCK:BMAL2-mediated transactivation. Together, CLOCK:BMAL1 and CLOCK:BMAL2 make additive contributions to PAI-1 gene transcription. While the abilities of these heterodimers to activate gene expression differ by twofold, the susceptibilities of these circadian activators to inhibition by period and cryptochrome proteins are equivalent and redox independent. Given that BMAL1 and BMAL2 differ in their spatiotemporal distributions, such distinctions may allow intrinsic circadian clocks to modulate the amplitudes of their oscillators, while maintaining circadian periodicity. In this way, fundamental circadian clock components may drive circadian variation in PAI-1, which in turn influences the pathogenesis, timing, and treatment of acute atherothrombotic events.
Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University Medical Center, 2220 Pierce Avenue, 383 PRB, Nashville, TN 37232, USA.
John A
JA
Schoenhard
Layton H
LH
Smith
Corrie A
CA
Painter
Mesut
M
Eren
Carl H
CH
Johnson
Douglas E
DE
Vaughan
D056930:ARNTL Transcription Factors; D000818:Animals; D001011:Aorta / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins; D056926:CLOCK Proteins; D002417:Cattle; D002940:Circadian Rhythm; D004730:Endothelium, Vascular / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
D056930:ARNTL Transcription Factors; D051792:Basic Helix-Loop-Helix Proteins; D056926:CLOCK Proteins; D017395:Plasminogen Activator Inhibitor 1; D015534:Trans-Activators; D014157:Transcription Factors; C538960:BMAL1 protein, human; C486615:BMAL2 protein, human; C538943:CLOCK protein, human