Rapid activation of the heat shock transcription factor, HSF1, by hypo-osmotic stress in mammalian cells.
Osmoregulation is important to living organisms for survival in responding to environmental changes of water and ionic strength. We demonstrated here for the first time that exposure of HeLa cells to a hypotonic medium (30% growth medium and 70% water) prominently induced the binding activity of the heat shock transcription factor (HSF). Pretreatment of cells with cycloheximide did not inhibit the induction of HSF-binding activity, indicating that the mechanisms of induction are independent of new protein synthesis. The magnitude of hypo-osmotic stress-induced HSF-binding activity was comparable with that induced by heat shock. The induction, as monitored by gel-mobility-shift assay, occurred within 5 min of hypo-osmotic stress and persisted at least up to 4 h in HeLa cells under the hypotonic conditions. Addition of sorbitol to the hypotonic medium abolished HSF activation. Hypo-osmotic stress-induced HSF binding could also be demonstrated in HeLa cells maintained in simple sorbitol solution by decreasing the sorbitol concentration from 300 mM to 200 mM or less. Competition analysis suggests that the effects of hypo-osmotic stress on HSF-binding activity was specific. Cross-linking experiments and Western-blot analysis demonstrated that hypo-osmotic stress induced trimerization of human heat shock factor 1 (HSF1) in intact HeLa cells, suggesting that trimer formation of HSF1 was responsible for inducing HSF-binding activity in hypo-osmotically stressed cells. However, unlike heat shock response, the activation of HSF by hypo-osmotic stress did not lead to accumulation of hsp70 mRNA in HeLa cells.
Huang
L E
LE
Department of Biological Sciences, Rutgers State University of New Jersey, Piscataway 08855-0939, USA.
Caruccio
L
L
Liu
A Y
AY
Chen
K Y
KY
D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D000076249:Heat Shock Transcription Factors; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009994:Osmolar Concentration; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D012996:Solutions; D013012:Sorbitol / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D018840:HSP70 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012996:Solutions; D014157:Transcription Factors; D013012:Sorbitol