title: Determination of the consensus binding sequence for the purified embryonic heat shock factor 2.
abstract: Heat shock transcription factors (HSFs) are characterized by their ability, upon activation, to bind to heat shock response elements (HSE) present in the promoter of their target genes. HSE are composed of inverted repeats of the pentamer nGAAm. In this study, we compare the embryonic HSF2 protein, purified from F9 embryonal carcinoma cells tumor, and the in vitro synthesized HSF2. We show that the context of HSF2 synthesis influences its thermosensitivity and DNA-binding properties. Therefore, we determined the consensus binding sequence for the purified embryonic HSF2 by the technique of systematic evolution of ligands by exponential enrichment (SELEX). We show that embryonic HSF2 prefers sites containing three or four nGAAm inverted pentamers and that its optimal binding sequence contains the 8-mer palindromic core 5'-TTCTAGAA-3'. The consensus binding sequence for the embryonic HSF2 will be very helpful to identify new targets for this factor, during developmental and differentiation processes.
authors.affiliation: Laboratoire de Biologie Moléculaire du Stress, Unité de Génétique Moléculaire UMR8541, Ecole Normale Supérieure, Paris, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Manuel
authors.forename: Murielle
authors.initials: M
authors.lastname: Rallu
authors.forename: Marie-Thérèse
authors.initials: MT
authors.lastname: Loones
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Zimarino
authors.forename: Valérie
authors.initials: V
authors.lastname: Mezger
authors.forename: Michel
authors.initials: M
authors.lastname: Morange
mesh_terms: D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism; D015723:Gene Library; D006360:Heat-Shock Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D051379:Mice; D011485:Protein Binding; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D006360:Heat-Shock Proteins; D014157:Transcription Factors; C070389:HSF2 protein, human; D004247:DNA