title: Activation of human period-1 by PKA or CLOCK/BMAL1 is conferred by separate signal transduction pathways.
abstract: Circadian clocks are self-sustained biochemical oscillators that autonomously generate a near-24 h cycle in the absence of external signals. The process of synchronization to the environment involves the transcriptional activation of several genes. Photic input signals from the retina are transduced via the retinohypothalamic tract to the central pacemaker located in the suprachiasmatic nuclei (SCN) of the hypothalamus. It is known that cells of peripheral organs possess similar molecular organizations, but the signal transductional pathways lack direct light entrainment. It has been assumed that the adaptation of peripheral organs to the SCN phase is achieved by the alternate usage of promoter elements. This question has been addressed by characterizing the signal transductional pathways regulating human Period-1 gene expression in human hepatoma cells (HuH-7). Plasmids coding for key modulators of circadian rhythm, hCLOCK, hBMAL1, and hCRY2 were used to analyze the activation of a human period-1 promoter luciferase (hPER1-luc) construct. Beside classical CLOCK/BMAL1 activation, hPER1-luc was also inducible by the overexpression of the catalytic subunit of PKA (Calpha). The cotransfection of dominant negative constructs to c-FOS, CREB, PKA, and C/EBP were used to characterize both regulatory pathways. It was found that hCLOCK/hBMAL1-mediated hPER1 activation was influenced by AP1, but not significantly by other regulators. Conversely, PKA-induced activation of hPER1 was reduced by the inhibition of CREB and the CCAAT-box binding protein C/EBP, but not by AP1. The present findings imply that CLOCK/BMAL1-mediated activation of hPER1 by AP1 and E-Box elements is distinct from peripheral transcriptional modulation via cAMP-induced CREB and C/EBP.
authors.affiliation: IPF PharmaCeuticals GmbH, Hannover, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Motzkus
authors.forename: Sabine
authors.initials: S
authors.lastname: Loumi
authors.forename: Christina
authors.initials: C
authors.lastname: Cadenas
authors.forename: Charles
authors.initials: C
authors.lastname: Vinson
authors.forename: Wolf-Georg
authors.initials: WG
authors.lastname: Forssmann
authors.forename: Erik
authors.initials: E
authors.lastname: Maronde
mesh_terms: D056930:ARNTL Transcription Factors; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D056926:CLOCK Proteins; D002460:Cell Line; D002940:Circadian Rhythm / Q000235:genetics* / Q000502:physiology*; D056931:Cryptochromes; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / Q000235:genetics / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D005420:Flavoproteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D056950:Period Circadian Proteins; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D056930:ARNTL Transcription Factors; D051792:Basic Helix-Loop-Helix Proteins; D056926:CLOCK Proteins; D056931:Cryptochromes; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits; D005136:Eye Proteins; D005420:Flavoproteins; D056950:Period Circadian Proteins; D011994:Recombinant Proteins; D015534:Trans-Activators; C538960:BMAL1 protein, human; C539022:PER1 protein, human; C538943:CLOCK protein, human; C463959:protein kinase A Calpha