The heat shock protein 90 antagonist geldanamycin alters chaperone association with p210bcr-abl and v-src proteins before their degradation by the proteasome.
Several important signaling proteins including transcription factors and protein kinases depend on heat shock protein (Hsp)-90 for stability. p210bcr-abl, a protein expressed in chronic myelogenous leukemia, is functionally inhibited by the benzoquinone ansamycin herbimycin A. Benzoquinone ansamycins also bind to and inhibit the activity of Hsp90. We now demonstrate that p210bcr-abl is complexed with Hsp90 and its cochaperone p23 in K562 chronic myelogenous leukemia cells. Brief exposure to the benzoquinone ansamycin Hsp90 inhibitor geldanamycin (GA) decreases the association of p210bcr-abl with Hsp90 and p23 and increases its association with the chaperones Hsp70 and p60Hop. GA has a similar effect on chaperone association with v-src, another Hsp90-dependent oncogenic kinase. Loss of Hsp90/p23 association and acquisition of Hsp70/p60Hop association of both p210bcr-abl and v-src precede GA-induced degradation of these kinases. GA-induced degradation is mediated by the proteasome because proteasome inhibitors block the effects of GA, causing both p210bcr-abl and v-src to accumulate in a detergent-insoluble cellular fraction. Both p210bcr-abl and v-src are more susceptible to GA-induced degradation than are their normal cellular counterparts, c-abl and c-src.
An
W G
WG
Department of Cell and Cancer Biology, Medicine Branch, National Cancer Institute, NIH, Rockville, Maryland 20850, USA.
Schulte
T W
TW
Neckers
L M
LM
D016475:3T3 Cells; D000818:Animals; D016227:Benzoquinones; D002457:Cell Extracts; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D016044:Fusion Proteins, bcr-abl / Q000378:metabolism*; D018922:HL-60 Cells; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D019746:Intramolecular Oxidoreductases; D020014:K562 Cells; D047029:Lactams, Macrocyclic; D046911:Macromolecular Substances; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism*; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D011233:Precipitin Tests; D000071876:Prostaglandin-E Synthases; D046988:Proteasome Endopeptidase Complex; D011487:Protein Conformation; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000378:metabolism; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000037:antagonists & inhibitors / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology*; D011980:Receptors, Progesterone / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
D016227:Benzoquinones; D002457:Cell Extracts; D004791:Enzyme Inhibitors; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D009097:Multienzyme Complexes; D010750:Phosphoproteins; D011809:Quinones; D011980:Receptors, Progesterone; D016044:Fusion Proteins, bcr-abl; D015688:Oncogene Protein pp60(v-src); D016315:Proto-Oncogene Proteins c-abl; D016392:Proto-Oncogene Proteins pp60(c-src); D019061:src-Family Kinases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D019746:Intramolecular Oxidoreductases; C490849:PTGES3 protein, human; D000071876:Prostaglandin-E Synthases; C001277:geldanamycin