Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones

Sonnet J H Arlander, Sara J. Felts, Jill M. Wagner, Bridget Stensgard, David O. Toft, Larry M Karnitz

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Checkpoint kinase 1 (Chk1), a serine/threonine kinase that regulates DNA damage checkpoints, is destabilized when heat shock protein 90 (Hsp90) is inhibited, suggesting that Chk1 is an Hsp90 client. In the present work we examined the interplay between Chk1 and Hsp90 in intact cells, identified a source of unchaperoned Chk1, and report the in vitro chaperoning of Chk1 in reticulocyte lysates and with purified chaperones and co-chaperones. We find that bacterially expressed Chk1 is post-translationally chaperoned to an active kinase. This reaction minimally requires Hsp90, Hsp70, Hsp40, Cdc37, and the protein kinase CK2. The co-chaperone Hop, although not essential for the activation of Chk1 in vitro, enhanced the chaperoning process, whereas the co-chaperone p23 did not stimulate the chaperoning reaction. Additionally, we found that the C-terminal regulatory domain of Chk1 affects the association of Chk1 with Hsp90. Collectively these results provide new insights into Hsp90-dependent chaperoning of a client kinase and identify a novel, biochemically tractable model system that will be useful to further dissect the Hsp90-dependent chaperoning of this important and ubiquitous class of Hsp90 clients.

Original languageEnglish (US)
Pages (from-to)2989-2998
Number of pages10
JournalJournal of Biological Chemistry
Volume281
Issue number5
DOIs
StatePublished - Feb 3 2006

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HSP90 Heat-Shock Proteins
Phosphotransferases
Checkpoint Kinase 1
Casein Kinase II
Humulus
Protein-Serine-Threonine Kinases
Reticulocytes
DNA Damage
Chemical activation
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. / Arlander, Sonnet J H; Felts, Sara J.; Wagner, Jill M.; Stensgard, Bridget; Toft, David O.; Karnitz, Larry M.

In: Journal of Biological Chemistry, Vol. 281, No. 5, 03.02.2006, p. 2989-2998.

Research output: Contribution to journalArticle

Arlander, Sonnet J H ; Felts, Sara J. ; Wagner, Jill M. ; Stensgard, Bridget ; Toft, David O. ; Karnitz, Larry M. / Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 5. pp. 2989-2998.
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