title: Deciphering the molecular mechanisms underlying the binding of the TWIST1/E12 complex to regulatory E-box sequences.
abstract: The TWIST1 bHLH transcription factor controls embryonic development and cancer processes. Although molecular and genetic analyses have provided a wealth of data on the role of bHLH transcription factors, very little is known on the molecular mechanisms underlying their binding affinity to the E-box sequence of the promoter. Here, we used an in silico model of the TWIST1/E12 (TE) heterocomplex and performed molecular dynamics (MD) simulations of its binding to specific (TE-box) and modified E-box sequences. We focused on (i) active E-box and inactive E-box sequences, on (ii) modified active E-box sequences, as well as on (iii) two box sequences with modified adjacent bases the AT- and TA-boxes. Our in silico models were supported by functional in vitro binding assays. This exploration highlighted the predominant role of protein side-chain residues, close to the heart of the complex, at anchoring the dimer to DNA sequences, and unveiled a shift towards adjacent ((-1) and (-1*)) bases and conserved bases of modified E-box sequences. In conclusion, our study provides proof of the predictive value of these MD simulations, which may contribute to the characterization of specific inhibitors by docking approaches, and their use in pharmacological therapies by blocking the tumoral TWIST1/E12 function in cancers.
authors.affiliation: Inserm UMR-S1052, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France LabEX DEVweCAN, Lyon, France UNIV UMR1052, Lyon 69008, France Centre Léon Bérard, Lyon 69373, France Université de Lyon1, ISPB, Lyon 69008, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Bouard
authors.affiliation: Université de Lyon1, ISPB, Lyon 69008, France Institut de Biochimie des protéines IBCP, Lyon 69007, France CNRS UMR 5305, Lyon 69007, France.
authors.forename: Raphael
authors.initials: R
authors.lastname: Terreux
authors.affiliation: Institut de Biochimie des protéines IBCP, Lyon 69007, France.
authors.forename: Mylène
authors.initials: M
authors.lastname: Honorat
authors.affiliation: Centre Léon Bérard, Lyon 69373, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Manship
authors.affiliation: CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France LabEX DEVweCAN, Lyon, France UNIV UMR1052, Lyon 69008, France Centre Léon Bérard, Lyon 69373, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Ansieau
authors.affiliation: Inserm UMR-S1052, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France LabEX DEVweCAN, Lyon, France UNIV UMR1052, Lyon 69008, France Centre Léon Bérard, Lyon 69373, France.
authors.forename: Arnaud M
authors.initials: AM
authors.lastname: Vigneron
authors.affiliation: Inserm UMR-S1052, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France LabEX DEVweCAN, Lyon, France UNIV UMR1052, Lyon 69008, France Centre Léon Bérard, Lyon 69373, France Université de Lyon1, ISPB, Lyon 69008, France Institut Universitaire de France, Paris 75231, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Puisieux
authors.affiliation: Inserm UMR-S1052, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon 69373, France LabEX DEVweCAN, Lyon, France UNIV UMR1052, Lyon 69008, France Centre Léon Bérard, Lyon 69373, France Université de Lyon1, ISPB, Lyon 69008, France Hospices Civils de Lyon, Laboratoire de Biochimie et Biologie Moléculaire du CHLS, Lyon 69003, France Lea.payen-gay@univ-lyon1.fr.
authors.forename: Léa
authors.initials: L
authors.lastname: Payen
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D024721:E-Box Elements; D006801:Humans; D006860:Hydrogen Bonding; D008958:Models, Molecular; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D058127:Transcription Factor 3 / Q000737:chemistry* / Q000378:metabolism; D051879:Twist-Related Protein 1 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D058127:Transcription Factor 3; D051879:Twist-Related Protein 1