Differentiation and fiber type-specific activity of a muscle creatine kinase intronic enhancer

• Published in: Skeletal muscle Vol. 1; no. 1; p. 25
• Main Authors: Tai, Phillip Wl, Fisher-Aylor, Katherine I, Himeda, Charis L, Smith, Catherine L, Mackenzie, Alexandra P, Helterline, Deri L, Angello, John C, Welikson, Robert E, Wold, Barbara J, Hauschka, Stephen D
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 07.07.2011; BioMed Central; BMC
• Subjects: Creatine kinase; Gene expression; Physiological aspects; Promoters (Genetics); United States
• ISSN: 2044-5040; 2044-5040
• DOI: 10.1186/2044-5040-1-25

Hundreds of genes, including muscle creatine kinase (MCK), are differentially expressed in fast- and slow-twitch muscle fibers, but the fiber type-specific regulatory mechanisms are not well understood. Modulatory region 1 (MR1) is a 1-kb regulatory region within MCK intron 1 that is highly active in terminally differentiating skeletal myocytes in vitro. A MCK small intronic enhancer (MCK-SIE) containing a paired E-box/myocyte enhancer factor 2 (MEF2) regulatory motif resides within MR1. The SIE's transcriptional activity equals that of the extensively characterized 206-bp MCK 5'-enhancer, but the MCK-SIE is flanked by regions that can repress its activity via the individual and combined effects of about 15 different but highly conserved 9- to 24-bp sequences. ChIP and ChIP-Seq analyses indicate that the SIE and the MCK 5'-enhancer are occupied by MyoD, myogenin and MEF2. Many other E-boxes located within or immediately adjacent to intron 1 are not occupied by MyoD or myogenin. Transgenic analysis of a 6.5-kb MCK genomic fragment containing the 5'-enhancer and proximal promoter plus the 3.2-kb intron 1, with and without MR1, indicates that MR1 is critical for MCK expression in slow- and intermediate-twitch muscle fibers (types I and IIa, respectively), but is not required for expression in fast-twitch muscle fibers (types IIb and IId). In this study, we discovered that MR1 is critical for MCK expression in slow- and intermediate-twitch muscle fibers and that MR1's positive transcriptional activity depends on a paired E-box MEF2 site motif within a SIE. This is the first study to delineate the DNA controls for MCK expression in different skeletal muscle fiber types.