title: Differential interaction of the dual alpha tropomyosin/N5 enhancer with multiple DNA binding proteins: N5 is a putative novel z-ZIP DNA binding protein.
abstract: The alpha tropomyosin (TM)/N5 enhancer is an SV40-like mammalian enhancer comprised of a 99 bp repeat with modular cis-acting regulatory elements exhibiting apparent hierarchical organization. The enhancer differentially regulates the alpha TM and N5 transcription units which exhibit distinct tissue-specific expression patterns and interacts with multiple myotube-associated nuclear DNA binding proteins that varied in size and amount. To further characterize the interaction with multiple myotube nuclear factors, comparative southwestern blot analyses were done with a panel of strategic DNA probes representative of modular enhancer sequences in the alpha TM/N5 enhancer and respective alpha TM and N5 promoter regions. Results demonstrate that multiple DNA binding proteins, which vary in size and amount, can interact with a particular enhancer modular sequence (delimited to 18 bp- to 38 bp-long); and that likewise, a DNA binding protein can bind specifically to different DNA enhancer modular sequences with apparent different affinities. Results also demonstrate DNA binding proteins that differentially bind to both enhancer modular sequences and respective promoter regions supporting a putative parsimonious mechanism for the approximation of enhancer and promoter elements as an alternative to the multi-protein stereospecific enhancer complex. Cogent to this interesting "head to head"/shared enhancer gene arrangement, we investigated the primary structure of the "other" transcription unit, N5. Nucleotide sequence analysis of the N5 cDNA reveals that it is a putative DNA binding protein representing a new structural class of transcription factors exhibiting a novel combinatorial motif: single zinc finger (DNA-binding)-leucine zipper (dimerization)--making it a z-ZIP instead of a b-ZIP (basic region/leucine zipper) protein.
authors.affiliation: Section of Molecular Genetics, Whitaker Cardiovascular Institute, Boston University School of Medicine, MA 02118.
authors.forename: N
authors.initials: N
authors.lastname: Ruiz-Opazo
authors.forename: J F
authors.initials: JF
authors.lastname: Cloix
authors.forename: V L
authors.initials: VL
authors.lastname: Herrera
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D016350:Leucine Zippers; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D011817:Rabbits; D014157:Transcription Factors; D014158:Transcription, Genetic; D014335:Tropomyosin / Q000235:genetics*; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors; D014335:Tropomyosin; D004247:DNA