A chemical switch for inhibitor-sensitive alleles of any protein kinase

Anthony C. Bishop, Jeffrey A. Ubersax, Dejah T. Pøtsch, Dina P. Matheos, Nathanael S. Gray, Justin Blethrow, Eiji Shimizu, Joseph Zhuo Tsien, Peter G. Schultz, Mark D. Rose, John L. Wood, David O. Morgan, Kevan M. Shokat

Research output: Contribution to journalArticle

743 Citations (Scopus)

Abstract

Protein kinases have proved to be largely resistant to the design of highly specific inhibitors, even with the aid of combinatorial chemistry1. The lack of these reagents has complicated efforts to assign specific signalling roles to individual kinases. Here we describe a chemical genetic strategy for sensitizing protein kinases to cell-permeable molecules that do not inhibit wild-type kinases2. From two inhibitor scaffolds, we have identified potent and selective inhibitors for sensitized kinases from five distinct subfamilies. Tyrosine and serine/threonine kinases are equally amenable to this approach. We have analysed a budding yeast strain carrying an inhibitor-sensitive form of the cyclindependent kinase Cdc28 (CDK1) in place of the wild-type protein. Specific inhibition of Cdc28 in vivo caused a pre-mitotic cell-cycle arrest that is distinct from the G1 arrest typically observed in temperature-sensitive cdc28 mutants3. The mutation that confers inhibitor-sensitivity is easily identifiable from primary sequence alignments. Thus, this approach can be used to systematically generate conditional alleles of protein kinases, allowing for rapid functional characterization of members of this important gene family.

Original languageEnglish (US)
Pages (from-to)395-401
Number of pages7
JournalNature
Volume407
Issue number6802
DOIs
StatePublished - Sep 21 2000
Externally publishedYes

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Protein Kinases
Phosphotransferases
Alleles
Saccharomycetales
Sequence Alignment
Protein-Serine-Threonine Kinases
Cell Cycle Checkpoints
Tyrosine
Mutation
Temperature
Genes
Proteins

ASJC Scopus subject areas

  • Medicine(all)
  • General

Cite this

Bishop, A. C., Ubersax, J. A., Pøtsch, D. T., Matheos, D. P., Gray, N. S., Blethrow, J., ... Shokat, K. M. (2000). A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature, 407(6802), 395-401. https://doi.org/10.1038/35030148

A chemical switch for inhibitor-sensitive alleles of any protein kinase. / Bishop, Anthony C.; Ubersax, Jeffrey A.; Pøtsch, Dejah T.; Matheos, Dina P.; Gray, Nathanael S.; Blethrow, Justin; Shimizu, Eiji; Tsien, Joseph Zhuo; Schultz, Peter G.; Rose, Mark D.; Wood, John L.; Morgan, David O.; Shokat, Kevan M.

In: Nature, Vol. 407, No. 6802, 21.09.2000, p. 395-401.

Research output: Contribution to journalArticle

Bishop, AC, Ubersax, JA, Pøtsch, DT, Matheos, DP, Gray, NS, Blethrow, J, Shimizu, E, Tsien, JZ, Schultz, PG, Rose, MD, Wood, JL, Morgan, DO & Shokat, KM 2000, 'A chemical switch for inhibitor-sensitive alleles of any protein kinase', Nature, vol. 407, no. 6802, pp. 395-401. https://doi.org/10.1038/35030148
Bishop AC, Ubersax JA, Pøtsch DT, Matheos DP, Gray NS, Blethrow J et al. A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature. 2000 Sep 21;407(6802):395-401. https://doi.org/10.1038/35030148
Bishop, Anthony C. ; Ubersax, Jeffrey A. ; Pøtsch, Dejah T. ; Matheos, Dina P. ; Gray, Nathanael S. ; Blethrow, Justin ; Shimizu, Eiji ; Tsien, Joseph Zhuo ; Schultz, Peter G. ; Rose, Mark D. ; Wood, John L. ; Morgan, David O. ; Shokat, Kevan M. / A chemical switch for inhibitor-sensitive alleles of any protein kinase. In: Nature. 2000 ; Vol. 407, No. 6802. pp. 395-401.
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