Identification of cardiomyocyte-specific transcriptional mechanism of c-myc

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We found that the expression of one gene is down-regulated in cardiomyocytes while up-regulated in HUVEC or smooth muscle cells by c-myc. To clarify this cardiomyocyte-specific transcriptional mechanism by c-myc, we developed the nuclear extraction method that can obtain structurally-intact transcription complexes binding with high-affinity to their specific DNA sequences. Next, we narrowed the range of putative binding site of c-myc transcription complexes in promoter region of the gene by using both comparative genomics approach and transactivation assay. We performed DNA affinity chromatography using oligonucleotides containing the binding site of c-myc transcription complex as bait and nuclear extracts from cardiomyocytes or HUVEC. Using a high-sensitive LC-MS/ MS technology, we identified cardiomyocyte-specific components in c-myc transcription complexes.