Heat-shock-induced denaturation of proteins. Characterization of the insolubilization of the interferon-induced p68 kinase.
Heat-shock stress causes inactivation and aggregation of various cellular proteins which become further insoluble. Previous studies have shown that the interferon-induced p68 kinase activity was greatly reduced in extracts of heat-shocked HeLa cells, and that the loss of activity was due to a decreased solubility of the enzyme. Here we show that the p68 kinase which is normally evenly distributed in the cytoplasm, aggregates as a thick ring around the nucleus in heat-shocked cells. The 70-kDa constitutive heat-shock proteins are major insolubilized proteins during stress and we find them to colocalize with the p68 kinase after stress. Treatments of cells with drugs which disrupt the cytoskeleton, such as colcemid and cytochalasin E, do not hinder the enzyme insolubilization during heat-shock. On the contrary, heat-protectors such as glycerol and deuterium oxide (D2O) keep the p68 kinase under a soluble and active form during heat-shock stress. Similarly, an attenuation of the insolubilization of this enzyme is observed in cells rendered thermo-tolerant by a previous heat-shock, suggesting that heat-shock proteins may also contribute to the protection. During the recovery period at normal temperature after heat-shock, resolubilization occurs and most of the enzyme is again recovered under an active soluble form.
Dubois
M F
MF
Groupe de Biologie Moléculaire du Stress Ecole Normale Supérieure, Paris, France.
Hovanessian
A G
AG
Bensaude
O
O
D015153:Blotting, Western; D003599:Cytoskeleton / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D011489:Protein Denaturation; D011494:Protein Kinases / Q000096:biosynthesis / Q000737:chemistry*; D019892:eIF-2 Kinase
D006360:Heat-Shock Proteins; D007372:Interferons; D011494:Protein Kinases; D019892:eIF-2 Kinase