Homeodomain protein MHox and MADS protein myocyte enhancer-binding factor-2 converge on a common element in the muscle creatine kinase enhancer

ABSTRACT Transcriptional regulatory element X (Trex) is a positive control site within the Muscle creatine kinase (MCK) enhancer. Cell culture and transgenic studies indicate that the Trex site is important for MCK expression in skeletal and cardiac muscle. After selectively enriching for the Trex-binding factor (TrexBF) using magnetic beads coupled to oligonucleotides containing either wild-type or mutant Trex sites, quantitative proteomics was used to identify TrexBF as Six4, a homeodomain transcription factor of the Six/sine oculis family, from a background of ∼900 copurifying proteins. Using gel shift assays and Six-specific antisera, we demonstrated that Six4 is TrexBF in mouse skeletal myocytes and embryonic day 10 chick skeletal and cardiac muscle, while Six5 is the major TrexBF in adult mouse heart. In cotransfection studies, Six4 transactivates the MCK enhancer as well as muscle-specific regulatory regions of Aldolase A and Cardiac troponin C via Trex/MEF3 sites. Our results are consistent with Six4 being a key regulator of muscle gene expression in adult skeletal muscle and in developing striated muscle. The Trex/MEF3 composite sequence ([C/A]ACC[C/T]GA) allowed us to identify novel putative Six-binding sites in six other muscle genes. Our proteomics strategy will be useful for identifying transcription factors from complex mixtures using only defined DNA fragments for purification.

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MHox: a mesodermally restricted homeodomain protein that binds an essential site in the muscle creatine kinase enhancer

Development ◽

10.1242/dev.115.4.1087 ◽

1992 ◽

Vol 115 (4) ◽

pp. 1087-1101 ◽

Cited By ~ 37

Author(s):

P. Cserjesi ◽

B. Lilly ◽

L. Bryson ◽

Y. Wang ◽

D.A. Sassoon ◽

...

Keyword(s):

Creatine Kinase ◽

Homeobox Genes ◽

Cell Types ◽

Limb Bud ◽

Homeodomain Protein ◽

Muscle Creatine Kinase ◽

Mesodermal Cell ◽

Adult Mice ◽

Mesodermal Origin ◽

Muscle Creatine

Myogenic helix-loop-helix (HLH) proteins, such as myogenin and MyoD, can activate muscle-specific transcription when introduced into a variety of nonmuscle cell types. Whereas cells of mesodermal origin are especially permissive to the actions of these myogenic regulators, many other cell types are refractory to myogenic conversion by them. Here we describe a novel homeodomain protein, MHox, that binds an A+T-rich element in the muscle creatine kinase (MCK) enhancer that is essential for muscle-specific transcription and trans-activation by myogenic HLH proteins. MHox is completely restricted to mesodermally derived cell types during embryogenesis and to established cell lines of mesodermal origin. In contrast to most other homeobox genes, MHox expression is excluded from the nervous system, with the highest levels observed in limb bud and visceral arches. In adult mice, MHox is expressed at high levels in skeletal muscle, heart and uterus. The DNA-binding properties and pattern of MHox expression are unique among homeobox genes and suggest a role for MHox as a transcriptional regulator that participates in the establishment of diverse mesodermal cell types.

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Complete cDNA-derived amino acid sequence of chick muscle creatine kinase.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)42538-5 ◽

1984 ◽

Vol 259 (24) ◽

pp. 15224-15227

Author(s):

C P Ordahl ◽

G L Evans ◽

T A Cooper ◽

G Kunz ◽

J C Perriard

Keyword(s):

Creatine Kinase ◽

Amino Acid ◽

Amino Acid Sequence ◽

Muscle Creatine Kinase ◽

Muscle Creatine

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Molecular docking and density functional theory studies on creatine, guanidinoacetic acid, and their phosphorylated analogues binding to muscle creatine kinase

Journal of Chemical Research ◽

10.1177/1747519820978583 ◽

2020 ◽

pp. 174751982097858

Author(s):

M Vraneš ◽

S Ostojić ◽

Č Podlipnik ◽

A Tot

Keyword(s):

Density Functional Theory ◽

Creatine Kinase ◽

Molecular Docking ◽

Density Functional ◽

Creatine Phosphate ◽

Docking Studies ◽

Muscle Creatine Kinase ◽

Functional Theory ◽

Guanidinoacetic Acid ◽

Muscle Creatine

Comparative molecular docking studies on creatine and guanidinoacetic acid, as well as their phosphorylated analogues, creatine phosphate, and phosphorylated guanidinoacetic acid, are investigated. Docking and density functional theory studies are carried out for muscle creatine kinase. The changes in the geometries of the ligands before and after binding to the enzyme are investigated to explain the better binding of guanidinoacetic acid and phosphorylated guanidinoacetic acid compared to creatine and creatine phosphate.

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Sodium barbital is a slow reversible inactivator of rabbit-muscle creatine kinase

Biochemistry and Cell Biology ◽

10.1139/o05-166 ◽

2006 ◽

Vol 84 (2) ◽

pp. 142-147

Author(s):

Feng Shi ◽

Tong-Jin Zhao ◽

Hua-Wei He ◽

Jie Li ◽

Xian-Gang Zeng ◽

...

Keyword(s):

Creatine Kinase ◽

Energy Homeostasis ◽

Inactivation Kinetics ◽

Rabbit Muscle ◽

Muscle Creatine Kinase ◽

Reversible Inactivation ◽

Inhibition Effect ◽

Sodium Barbital ◽

Muscle Creatine ◽

Detailed Kinetics

As a depressant of the central nervous system, the clinical effect of sodium barbital has been extensively studied. Here we report on sodium barbital as an inhibitor of rabbit-muscle creatine kinase (CK), which plays a significant role in energy homeostasis in the muscles. Although sodium barbital gradually inhibits the activity of CK with increased concentration, the inhibition effect can be completely reversed by dilution, indicating that the inactivation process is reversible. Detailed kinetics analysis, according to a previously presented theory, indicates that sodium barbital functions as a non complexing inhibitor, and its inhibition effect on CK is a slow reversible inactivation. In this study, a kinetic model of the substrate reaction is presented, and the microscopic rate constants for the reaction of sodium barbital with the free enzyme and the enzyme–substrate complexes are determined. Kinetic analysis reveals that sodium barbital might compete with both creatine and ATP, but mainly with creatine, to inhibit the activity of CK. The results suggest that CK might be a target for sodium barbital in vivo.Key words: creatine kinase; inactivation; kinetics; sodium barbital.

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Treatment of human muscle creatine kinase with glutaraldehyde preferentially increases the immunogenicity of the native conformation and permits production of high-affinity monoclonal antibodies which recognize two distinct surface epitopes

Journal of Immunological Methods ◽

10.1016/0022-1759(89)90100-2 ◽

1989 ◽

Vol 125 (1-2) ◽

pp. 251-259 ◽

Cited By ~ 15

Author(s):

Nguyen thi Man ◽

Alison J. Cartwright ◽

Karen M. Andrews ◽

Glenn E. Morris

Keyword(s):

Creatine Kinase ◽

Monoclonal Antibodies ◽

Human Muscle ◽

Native Conformation ◽

Muscle Creatine Kinase ◽

High Affinity ◽

Muscle Creatine

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Brain and muscle creatine kinase genes contain common TA-rich recognition protein-binding regulatory elements.

Molecular and Cellular Biology ◽

10.1128/mcb.10.9.4826 ◽

1990 ◽

Vol 10 (9) ◽

pp. 4826-4836 ◽

Cited By ~ 29

Author(s):

R A Horlick ◽

G M Hobson ◽

J H Patterson ◽

M T Mitchell ◽

P A Benfield

Keyword(s):

Creatine Kinase ◽

Binding Site ◽

Regulatory Element ◽

Gene Promoter ◽

Regulatory Elements ◽

Muscle Creatine Kinase ◽

Enhancer Activity ◽

L6 Myotubes ◽

Recognition Protein ◽

Muscle Creatine

We have previously reported that the rat brain creatine kinase (ckb) gene promoter contains an AT-rich sequence that is a binding site for a protein called TARP (TA-rich recognition protein). This AT-rich segment is a positively acting regulatory element for the ckb promoter. A similar AT-rich DNA segment is found at the 3' end of the 5' muscle-specific enhancer of the rat muscle creatine kinase (ckm) gene and has been shown to be necessary for full muscle-specific enhancer activity. In this report, we show that TARP binds not only to the ckb promoter but also to the AT-rich segment at the 3' end of the muscle-specific ckm enhancer. A second, weaker TARP-binding site was identified in the ckm enhancer and lies at the 5' end of the minimal enhancer segment. TARP was found in both muscle cells (C2 and L6 myotubes) and nonmuscle (HeLa) cells and appeared to be indistinguishable from both sources, as judged by gel retardation and footprinting assays. The TARP-binding sites in the ckm enhancer and the ckb promoter were found to be functionally interchangeable. We propose that TARP is active in both muscle and nonmuscle cells and that it is one of many potential activators that may interact with muscle-specific regulators to determine the myogenic phenotype.

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