Binding of myc proteins to canonical and noncanonical DNA sequences.
Using an in vitro binding-site selection assay, we have demonstrated that c-Myc-Max complexes bind not only to canonical CACGTG or CATGTG motifs that are flanked by variable sequences but also to noncanonical sites that consist of an internal CG or TG dinucleotide in the context of particular variations in the CA--TG consensus. None of the selected sites contain an internal TA dinucleotide, suggesting that Myc proteins necessarily bind asymmetrically in the context of a CAT half-site. The noncanonical sites can all be bound by proteins of the Myc-Max family but not necessarily by the related CACGTG- and CATGTG-binding proteins USF and TFE3. Substitution of an arginine that is conserved in these proteins into MyoD (MyoD-R) changes its binding specificity so that it recognizes CACGTG instead of the MyoD cognate sequence (CAGCTG). However, like USF and TFE3, MyoD-R does not bind to all of the noncanonical c-Myc-Max sites. Although this R substitution changes the internal dinucleotide specificity of MyoD, it does not significantly alter its wild-type binding sequence preferences at positions outside of the CA--TG motif, suggesting that it does not dramatically change other important amino acid-DNA contacts; this observation has important implications for models of basic-helix-loop-helix protein-DNA binding.
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
T K
TK
Blackwell
J
J
Huang
A
A
Ma
L
L
Kretzner
F W
FW
Alt
R N
RN
Eisenman
H
H
Weintraub
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors; D001665:Binding Sites; D001667:Binding, Competitive; D004268:DNA-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000378:metabolism; D017570:MyoD Protein; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014157:Transcription Factors
D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D009124:Muscle Proteins; C067850:Myc associated factor X; D017570:MyoD Protein; D009838:Oligodeoxyribonucleotides; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors