Geldanamycin induces heat shock protein expression through activation of HSF1 in K562 erythroleukemic cells.
HSP90 is one of the most abundant heat shock proteins (HSPs) in eukaryotic cells and is found in complex with several regulatory proteins such as kinases and transcription factors. Geldanamycin (GA), a benzoquinone ansamycin, specifically binds to HSP90 and disrupts the interaction of HSP90 and target proteins. Thus, GA has been used as a specific inhibitor of HSP90. In this study, we examined whether GA could affect protein synthesis and gene expression in the human erythroleukemic cell line K562. Treatment with GA, but not herbimycin A (another benzoquinone ansamycin), highly induced a 70-kDa protein, which was revealed to be HSP70 by immunoblotting and immunoprecipitation with anti-HSP70 antibody. The expression of HSP28 was also enhanced by GA. Furthermore, GA induced the activation of heat shock factor 1 (HSF1), but not HSF2, as determined by electromobility shift and electromobility supershift assay. In addition, similar to heat shock treatment, GA induced the phosphorylation of HSF1. Heat shock element-binding activity and phosphorylation of HSF1 were attenuated 3 h after GA treatment. These results indicate that the functional inactivation of HSP90 by GA potentially stimulates the expression of heat shock proteins through activation of HSF1.
Kim
H R
HR
Department of Molecular Biology, College of Natural Sciences, Pusan National University, Korea.
Kang
H S
HS
Kim
H D
HD
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D016227:Benzoquinones; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D018841:HSP90 Heat-Shock Proteins / Q000096:biosynthesis; D000076249:Heat Shock Transcription Factors; D006801:Humans; D020014:K562 Cells; D047029:Lactams, Macrocyclic; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D011233:Precipitin Tests; D011809:Quinones / Q000494:pharmacology*; D014157:Transcription Factors
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D047029:Lactams, Macrocyclic; D011809:Quinones; D014157:Transcription Factors; C001277:geldanamycin