title: Heat-induced chaperone activity of HSP90.
abstract: The 90-kDa stress protein, HSP90, is a major cytosolic protein ubiquitously distributed in all species. Using two substrate proteins, dihydrofolate reductase (DHFR) and firefly luciferase, we demonstrate here that HSP90 newly acquires a chaperone activity when incubated at temperatures higher than 46 degrees C, which is coupled with self-oligomerization of HSP90. While chemically denatured DHFR refolds spontaneously upon dilution from denaturant, oligomerized HSP90 bound DHFR during the process of refolding and prevented it from renaturation. DHFR was released from the complex with HSP90 by incubating with GroEL/ES complexes in an ATP-dependent manner and refolded into the native form. alpha-Casein inhibited the binding of DHFR to HSP90 and also chased DHFR from the complex with HSP90. These results suggest that HSP90 binds substrates to maintain them in a folding-competent structure. Furthermore, we found that HSP90 prevents luciferase from irreversible thermal denaturation and enables it to refold when postincubated with reticulocyte lysates. This heat-induced chaperone activity of HSP90 associated with its oligomerization may have a pivotal role in protection of cells from thermal damages.
authors.affiliation: Department of Cell Biology, Tokyo Metropolitan Institute of Medical Science, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Yonehara
authors.forename: Y
authors.initials: Y
authors.lastname: Minami
authors.forename: Y
authors.initials: Y
authors.lastname: Kawata
authors.forename: J
authors.initials: J
authors.lastname: Nagai
authors.forename: I
authors.initials: I
authors.lastname: Yahara
mesh_terms: D000818:Animals; D001483:Base Sequence; D018833:Chaperonins / Q000378:metabolism*; D017931:DNA Primers; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D013552:Swine; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry / Q000378:metabolism
chemical_list: D017931:DNA Primers; D018841:HSP90 Heat-Shock Proteins; D008156:Luciferases; D013762:Tetrahydrofolate Dehydrogenase; D018833:Chaperonins