The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone

Monica G. Marcu, Ahmed Chadli, Ilham Bouhouche, Maria Catelli, Leonard M. Neckers

Research output: Contribution to journalArticle

397 Citations (Scopus)

Abstract

Heat shock protein 90 (Hsp90), one of the most abundant chaperones in eukaryotes, participates in folding and stabilization of signal-transducing molecules including steroid hormone receptors and protein kinases. The amino terminus of Hsp90 contains a non-conventional nucleotide-binding site, related to the ATP-binding motif of bacterial DNA gyrase. The anti-tumor agents geldanamycin and radicicol bind specifically at this site and induce destabilization of Hsp90-dependent client proteins. We recently demonstrated that the gyrase inhibitor novobiocin also interacts with Hsp90, altering the affinity of the chaperone for geldanamycin and radicicol and causing in vitro and in vivo depletion of key regulatory Hsp90-dependent kinases including v-Src, Raf-1, and p185(ErbB2). In the present study we used deletion/mutation analysis to identify the site of interaction of novobiocin with Hsp90, and we demonstrate that the novobiocin-binding site resides in the carboxyl terminus of the chaperone. Surprisingly, this motif also recognizes ATP, and ATP and novobiocin efficiently compete with each other for binding to this region of Hsp90. Novobiocin interferes with association of the cochaperones Hsc70 and p23 with Hsp90. These results identify a second site on Hsp90 where the binding of small molecule inhibitors can significantly impact the function of this chaperone, and they support the hypothesis that both amino- and carboxyl-terminal domains of Hsp90 interact to modulate chaperone activity.

Original languageEnglish (US)
Pages (from-to)37181-37186
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number47
DOIs
StatePublished - Nov 24 2000

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Novobiocin
HSP90 Heat-Shock Proteins
Adenosine Triphosphate
Steroid hormones
Binding Sites
DNA Gyrase
Bacterial DNA
Molecules
Steroid Receptors
Sequence Deletion
Eukaryota
Protein Binding
Protein Kinases
Tumors
Phosphotransferases
Nucleotides
Stabilization
Hormones

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone. / Marcu, Monica G.; Chadli, Ahmed; Bouhouche, Ilham; Catelli, Maria; Neckers, Leonard M.

In: Journal of Biological Chemistry, Vol. 275, No. 47, 24.11.2000, p. 37181-37186.

Research output: Contribution to journalArticle

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