Anomalous interaction of Sp1 and specific binding of an E‐box‐binding protein with the regulatory elements of the Na,K‐ATPase α2 subunit gene promoter
- 1 November 1993
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 218 (1) , 195-204
- https://doi.org/10.1111/j.1432-1033.1993.tb18365.x
Abstract
We identified cis elements in the 5'-flanking region of rat Na,K-ATPase alpha 2 subunit gene (Atp1a2) using transient transfection assays in L6 rat skeletal muscle myoblast cells. By 5'-deletion mutation analysis, the region between nucleotide positions -175 and -108 was identified as a positive regulatory region. In the region, the distal E box (nucleotides -144 to -139) acts as a negative regulatory element, and the Sp1 consensus sequence (nucleotides -123 to -118) and the GGGAGG sequence (nucleotides -114 to -109) act as positive regulatory elements. Gel-retardation analysis revealed that binding factors are an E-box-binding protein and Sp1. DNase I footprinting and methylation-interference analyses revealed that Sp1 binds to the region from nucleotides -122 to -101 and the E-box-binding protein to the region from nucleotides -144 to -136. T4 DNA polymerase footprinting revealed that there are three Sp1-binding sites in the region and that Sp1 binds to one of the three sites in a mutually exclusive manner. The mechanism by which Sp1 activates the Atp1a2 promoter is discussed.Keywords
This publication has 37 references indexed in Scilit:
- Sp1 binds to the adhesion molecule on glia regulatory element that functions as a positive transcription regulatory element in astrocytesJournal of Neuroscience Research, 1993
- Synergism in transcriptional activation: a kinetic view.Genes & Development, 1993
- Oncogenic germ-line mutations in Sp1 and ATF sites in the human retinoblastoma geneNature, 1991
- An inhibitory domain of E12 transcription factor prevents DNA binding in E12 homodimers but not in E12 heterodimersCell, 1991
- Differences and Similarities in DNA-Binding Preferences of MyoD and E2A Protein Complexes Revealed by Binding Site SelectionScience, 1990
- GC box binding induces phosphorylation of Sp1 by a DNA-dependent protein kinasePublished by Elsevier ,1990
- The MyoD DNA binding domain contains a recognition code for muscle-specific gene activationCell, 1990
- O-glycosylation of eukaryotic transcription factors: Implications for mechanisms of transcriptional regulationCell, 1988
- Molecular cloning of three distinct forms of the Na+,K+-ATPase .alpha.-subunit from rat brainBiochemistry, 1986
- Purification and Biochemical Characterization of the Promoter-Specific Transcription Factor, Sp1Science, 1986