Sequences flanking the E-box contribute to cooperative binding by c-Myc/Max heterodimers to adjacent binding sites
Program in Gene Function and Expression; Program in Molecular Medicine
Animals; Base Sequence; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Basic-Leucine Zipper Transcription Factors; Binding Sites; DNA-Binding Proteins; Helix-Loop-Helix Motifs; Humans; Ornithine Decarboxylase; *Promoter Regions, Genetic; Protein Precursors; Proto-Oncogene Proteins c-myc; Rats; Thymosin; *Transcription Factors
Genetics and Genomics
Previously, we have shown that c-Myc/Max heterodimers, bind cooperatively to the two adjacent, canonical E-boxes (CACGTG) located in the rat ornithine decarboxylase (ODC) gene. In order to study this in more detail, we changed the length of the linker that separates the two E-boxes, as well as their flanking sequences. We found that high affinity, cooperative binding requires a minimal linker length of 1-4 bp and that the binding affinity is influenced by E-box flanking sequences. Binding to the c-Myc responsive element of prothymosin alpha, containing both a canonical and a noncanonical E-box (CAAGTG) was also studied. As shown by DNAseI footprinting analysis, only the canonical E-box is bound by c-Myc/Max and c-Max/Max dimers. Replacing the noncanonical site with a canonical E-box only partially restored high affinity, cooperative binding. By making hybrid fragments between ODC and prothymosin alpha, we found that nucleotides in the linker between the E-boxes influence the affinity of c-Myc/Max heterodimers. Taken together, our results show that E-box sequences and sequences in the linker separating both E-boxes influence cooperative, high affinity binding by c-Myc/Max dimers.
DOI of Published Version
Biochim Biophys Acta. 1998 Apr 29;1397(2):189-201. Link to article on publisher's site
Biochimica et biophysica acta
Walhout, Albertha J. M.; van der Vliet, P. C.; and Timmers, H. Th. M., "Sequences flanking the E-box contribute to cooperative binding by c-Myc/Max heterodimers to adjacent binding sites" (1998). Program in Gene Function and Expression Publications and Presentations. 8.