Alternative multimeric structures affect myogenin DNA binding activity.
The native molecular weight of the basic helix-loop-helix (bHLH) proteins myogenin, MyoD, and E12 was calculated from their mobilities on sucrose gradients and molecular sieve chromatography. The muscle bHLH proteins associate to form a variety of higher order complexes, most of which are larger than dimers. Homodimers bind to DNA sequences such as the MEF-1 site in the creatine kinase enhancer whereas homotetramers and larger forms do not recognize this DNA sequence. The ubiquitous bHLH protein E12 forms monomers or homodimers with little evidence for higher order complexes. Mixtures of myogenin and E12 show some heterodimeric structures, but most of the myogenin remains in large complexes. This result using purified proteins is also obtained in nuclear extracts from differentiated myotubes, in which most of the myogenin is present in large complexes that do not bind to the creatine kinase enhancer. A fusion protein containing only the myogenin HLH region forms large homomeric complexes. A model is presented in which each helix associates with a different subunit to form chains or ring structures to explain these observations. The partition of myogenin in nuclear extracts into dimers that recognize known DNA sequences and higher order complexes that do not raises important new issues concerning the regulation of skeletal muscle bHLH protein activity during myogenesis.
Farmer
K
K
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235.
Catala
F
F
Wright
W E
WE
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D046911:Macromolecular Substances; D008961:Models, Structural; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism; D017570:MyoD Protein; D018008:Myogenin; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013379:Substrate Specificity; D015534:Trans-Activators / Q000378:metabolism
D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D009124:Muscle Proteins; D017570:MyoD Protein; D018008:Myogenin; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D004247:DNA; D005982:Glutathione Transferase