title: Negative regulation of beta enolase gene transcription in embryonic muscle is dependent upon a zinc finger factor that binds to the G-rich box within the muscle-specific enhancer.
abstract: We have previously identified a muscle-specific enhancer within the first intron of the human beta enolase gene. Present in this enhancer are an A/T-rich box that binds MEF-2 protein(s) and a G-rich box (AGTGGGGGAGGGGGCTGCG) that interacts with ubiquitously expressed factors. Both elements are required for tissue-specific expression of the gene in skeletal muscle cells. Here, we report the identification and characterization of a Kruppel-like zinc finger protein, termed beta enolase repressor factor 1, that binds in a sequence-specific manner to the G-rich box and functions as a repressor of the beta enolase gene transcription in transient transfection assays. Using fusion polypeptides of beta enolase repressor factor 1 and the yeast GAL4 DNA-binding domain, we have identified an amino-terminal region responsible for the transcriptional repression activity, whereas a carboxyl-terminal region was shown to contain a potential transcriptional activation domain. The expression of this protein decreases in developing skeletal muscles, correlating with lack of binding activity in nuclear extract from adult skeletal tissue, in which novel binding activities have been detected. These results suggest that in addition to the identified factor, which functionally acts as a negative regulator and is enriched in embryonic muscle, the G-rich box binds other factors, presumably exerting a positive control on transcription. The interplay between factors that repress or activate transcription may constitute a developmentally regulated mechanism that modulates beta enolase gene expression in skeletal muscle.
authors.affiliation: Istituto di Biologia dello Sviluppo del Consiglio Nazionale delle Ricerche, Via Ugo La Malfa 153, 90146 Palermo, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Passantino
authors.forename: V
authors.initials: V
authors.lastname: Antona
authors.forename: G
authors.initials: G
authors.lastname: Barbieri
authors.forename: P
authors.initials: P
authors.lastname: Rubino
authors.forename: R
authors.initials: R
authors.lastname: Melchionna
authors.forename: G
authors.initials: G
authors.lastname: Cossu
authors.forename: S
authors.initials: S
authors.lastname: Feo
authors.forename: A
authors.initials: A
authors.lastname: Giallongo
mesh_terms: D000375:Aging / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004742:Enhancer Elements, Genetic; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000196:embryology / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D010751:Phosphopyruvate Hydratase