Properties and partial characterization of the heat-shock factor from Tetrahymena pyriformis.
A heat-shock-factor-binding activity was identified in Tetrahymena pyriformis whole-cell extracts and was further purified by sequential heparin-agarose and sequence-specific oligonucleotide affinity chromatography. Tetrahymena heat-shock factor (HSF) was able to bind to the heat-shock elements (HSE) both before and after thermal stress, although heat shock altered both the HSE-binding affinity and the protein.DNA-complex mobility on polyacrylamide gels. The mobility difference was significantly reduced by treatment of the proteins with phosphatase. The HSE-binding proteins, isolated by oligonucleotide-affinity chromatography, migrated on SDS/polyacrylamide gels as a closely spaced doublet to about 70 kDa. Polypeptides with similar molecular mass were recovered from preparative band-shift gels indicating that both are components of the protein.DNA complex.
Avides
M do C
Mdo C
Laboratório de Bioquimica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Portugal.
Sunkel
C E
CE
Moradas-Ferreira
P
P
Rodrigues-Pousada
C
C
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D016172:DNA Fingerprinting; D004268:DNA-Binding Proteins / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010957:Plasmids; D013769:Tetrahymena pyriformis / Q000378:metabolism*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins