Open AccessDetermination of the consensus binding sequence for the purified embryonic heat shock factor 2
Author(s) -
Manuel Martine,
Rallu Murielle,
Loones MarieThérèse,
Zimarino Vincenzo,
Mezger Valérie,
Morange Michel
Publication year - 2002
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1033.2002.02917.x
Subject(s) - pentamer , consensus sequence , heat shock factor , systematic evolution of ligands by exponential enrichment , embryonic stem cell , biology , microbiology and biotechnology , transcription factor , context (archaeology) , gene , genetics , heat shock protein , peptide sequence , biochemistry , rna , hsp70 , paleontology
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.