Cell Stress and Chaperones

, Volume 19, Issue 5, pp 695–703

Cdc37 engages in stable, S14A mutation-reinforced association with the most atypical member of the yeast kinome, Cdk-activating kinase (Cak1)

  • Stefan Millson
  • Patricija van Oosten-Hawle
  • Mohammed A. Alkuriji
  • Andrew Truman
  • Marco Siderius
  • Peter W. Piper
Original Paper

DOI: 10.1007/s12192-014-0497-4

Cite this article as:
Millson, S., van Oosten-Hawle, P., Alkuriji, M.A. et al. Cell Stress and Chaperones (2014) 19: 695. doi:10.1007/s12192-014-0497-4
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Abstract

In most eukaryotes, Cdc37 is an essential chaperone, transiently associating with newly synthesised protein kinases in order to promote their stabilisation and activation. To determine whether the yeast Cdc37 participates in any stable protein interactions in vivo, genomic two-hybrid screens were conducted using baits that are functional as they preserve the integrity of the conserved N-terminal region of Cdc37, namely a Cdc37-Gal4 DNA binding domain (BD) fusion in both its wild type and its S14 nonphosphorylatable (Cdc37(S14A)) mutant forms. While this failed to identify the protein kinases previously identified as Cdc37 interactors in pull-down experiments, it did reveal Cdc37 engaging in a stable association with the most atypical member of the yeast kinome, cyclin-dependent kinase (Cdk1)-activating kinase (Cak1). Phosphorylation of the conserved S14 of Cdc37 is normally crucial for the interaction with, and stabilisation of, those protein kinase targets of Cdc37, Cak1 is unusual in that the lack of this Cdc37 S14 phosphorylation both reinforces Cak1:Cdc37 interaction and does not compromise Cak1 expression in vivo. Thus, this is the first Cdc37 client kinase found to be excluded from S14 phosphorylation-dependent interaction. The unusual stability of this Cak1:Cdc37 association may partly reflect unique structural features of the fungal Cak1.

Keywords

Yeast two-hybridCdc37Molecular chaperoneCak1

Copyright information

© Cell Stress Society International 2014

Authors and Affiliations

  • Stefan Millson
    • 1
  • Patricija van Oosten-Hawle
    • 2
    • 5
  • Mohammed A. Alkuriji
    • 1
  • Andrew Truman
    • 3
  • Marco Siderius
    • 4
  • Peter W. Piper
    • 1
  1. 1.Department of Molecular Biology and BiotechnologyUniversity of SheffieldSheffieldUK
  2. 2.Department of Biochemistry and Molecular Biology, Faculty of ScienceVU UniversityAmsterdamThe Netherlands
  3. 3.University of ChicagoChicagoUSA
  4. 4.Department of Medicinal Chemistry, Faculty of ScienceVU UniversityAmsterdamThe Netherlands
  5. 5.Department of Biochemistry, Molecular Biology and Cell BiologyNorthwestern UniversityEvanstonUSA