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Circadian rhythms are important for healthy cardiovascular physiology and are regulated at the molecular level by a circadian clock mechanism. We and others previously demonstrated that 9–13% of the cardiac transcriptome is rhythmic over 24 h daily cycles; the heart is genetically a different organ day versus night. However, which rhythmic mRNAs are regulated by the circadian mechanism is not known. Here, we used open access bioinformatics databases to identify 94 transcripts with expression profiles characteristic of CLOCK and BMAL1 targeted genes, using the CircaDB website and JTK_Cycle. Moreover, 22 were highly expressed in the heart as determined by the BioGPS website. Furthermore, 5 heart-enriched genes had human/mouse conserved CLOCK:BMAL1 promoter binding sites (E-boxes), as determined by UCSC table browser, circadian mammalian promoter/enhancer database PEDB, and the European Bioinformatics Institute alignment tool (EMBOSS). Lastly, we validated findings by demonstrating that  Titin cap  ( Tcap ,  telethonin ) was targeted by transcriptional activators CLOCK and BMAL1 by showing 1)  Tcap  mRNA and TCAP protein had a diurnal rhythm in murine heart; 2) cardiac  Tcap  mRNA was rhythmic in animals kept in constant darkness; 3)  Tcap  and control  Per2  mRNA expression and cyclic amplitude were blunted in Clock Δ19/Δ19  hearts; 4) BMAL1 bound to the  Tcap  promoter by ChIP assay; 5) BMAL1 bound to  Tcap  promoter E-boxes by biotinylated oligonucleotide assay; and 6) CLOCK and BMAL1 induced  tcap  expression by luciferase reporter assay. Thus this study identifies circadian regulated genes  in silico , with validation of  Tcap,  a critical regulator of cardiac Z-disc sarcomeric structure and function.

plos one

Circadian Regulation of Myocardial Sarcomeric Titin-cap (Tcap, Telethonin): Identification of Cardiac Clock-Controlled Genes Using Open Access Bioinformatics Data