The molecular chaperones Hsp90 and Hsc70 are both necessary and sufficient to activate hormone binding by glucocorticoid receptor.
Glucocorticoid receptors must be complexed with Hsp90 in order to bind steroids, and it has been reported that at least three other proteins, Hop, Hsc70, and a J-domain protein (either Hsp40 or Ydj1), are required for formation of active Hsp90-steroid receptor complex. In the present study, we reinvestigated activation of stripped steroid receptors isolated from either L cells or WCL2 cells. Surprisingly, we found, using highly purified proteins, that only Hsp90 and Hsc70 are required for the activation of glucocorticoid receptors in the presence of steroids; in the absence of steroids, either p23 or molybdate are also required as reported previously. Addition of Hop or Ydj1 had no affect on the rate or magnitude of the activation of the stripped receptors, and quantitative Western blots confirmed that neither Hop or Hsp40 were present in our protein preparations or in the stripped receptors. Furthermore, a truncated recombinant Hsp70 that does not bind Hop or Hsp40 was as effective as wild-type Hsp70 in activating stripped receptor. Since Hsc70 does not bind directly to Hsp90 but both proteins bind to Hop, it has been suggested that Hop acts as a bridge between Hsp90 and Hsp70. However, we found that after Hsc70 or Hsp90 bind directly to the stripped receptors, they are fully reactivated by Hsp90 or Hsc70, respectively. We, therefore, conclude that Hsp90 and Hsc70 bind independently to stripped glucocorticoid receptors and alone are sufficient to activate them to bind steroids.
Rajapandi
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Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Greene
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LE
Eisenberg
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D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D005938:Glucocorticoids / Q000378:metabolism*; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D015398:Signal Transduction
D002352:Carrier Proteins; D005938:Glucocorticoids; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; C494372:Hspa8 protein, mouse; D018832:Molecular Chaperones; D011965:Receptors, Glucocorticoid