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New structural determinants for c‐Myc specific heterodimerization with Max and development of a novel homodimeric c‐Myc b‐HLH‐LZ ,
Author(s) -
Beaulieu MarieEve,
McDuff FrançoisOlivier,
Frappier Vincent,
Montagne Martin,
Naud JeanFrançois,
Lavigne Pierre
Publication year - 2012
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.2203
Subject(s) - basic helix loop helix , transcription factor , leucine zipper , mutant , chemistry , psychological repression , stereochemistry , dna binding protein , biochemistry , biology , gene , gene expression
c‐Myc must heterodimerize with Max to accomplish its functions as a transcription factor. This specific heterodimerization occurs through the b‐HLH‐LZ (basic region, helix 1‐loop‐helix 2‐leucine zipper) domains. In fact, many studies have shown that the c‐Myc b‐HLH‐LZ (c‐Myc'SH) preferentially forms a heterodimer with the Max b‐HLH‐LZ (Max'SH). The primary mechanism underlying the specific heterodimerization lies on the destabilization of both homodimers and the formation of a more stable heterodimer. In this regard, it has been widely reported that c‐Myc'SH has low solubility and homodimerizes poorly and that repulsions within the LZ domain account for the homodimer instability. Here, we show that replacing one residue in the basic region and one residue in Helix 1 (H 1 ) of c‐Myc'SH with corresponding residues conserved in b‐HLH proteins confers to c‐Myc'SH a higher propensity to form a stable homodimer in solution. In stark contrast to the wild‐type protein, this double mutant (L362R, R367L) of the c‐Myc b‐HLH‐LZ (c‐Myc'RL) shows limited heterodimerization with Max'SH in vitro . In addition, c‐Myc'RL forms highly stable and soluble complexes with canonical as well as non‐canonical E‐box probes. Altogether, our results demonstrate for the first time that structural determinants driving the specific heterodimerization of c‐Myc and Max are embedded in the basic region and H 1 of c‐Myc and that these can be exploited to engineer a novel homodimeric c‐Myc b‐HLH‐LZ with the ability of binding the E‐box sequence autonomously and with high affinity. Copyright © 2012 John Wiley & Sons, Ltd.