Protein Phosphatase Protocols

Volume 365 of the series Methods in Molecular Biology pp 309-322

Targeting of PP2C in Budding Yeast

  • Irene M. OtaAffiliated withHuntsman Cancer Institute, University of Utah
  • , James MapesAffiliated withDepartment of Molecular, Cellular, and Developmental Biology, University of Colorado

* Final gross prices may vary according to local VAT.

Get Access


Type 2C Ser/Thr phosphatases or PP2Cs are monomeric metal-requiring protein phosphatases that are present in prokaryotes and eukaryotes. In the yeast Saccharomyces cerevisiae, there are seven PP2Cs called PTCs (phosphatase 2C). Molecular genetic studies have implicated PTCs in many different functions, including RNA splicing, the unfolded protein response, mitogen-activated protein kinase (MAPK) pathway, and cell-cycle regulation. We have shown that three PTCs (Ptc1, Ptc2, and Ptc3), regulate the stress-activated high-osmolarity glycerol (HOG) mitogen-activated protein kinase (MAPK) pathway. Proteomics studies have provided additional possible functions for these phosphatases by identifying interacting proteins. These studies have also provided the possible means by which these phosphatases are targeted to their substrates. For example, Nbp2-Ptc1 was identified as an interacting pair in yeast two-hybrid studies, and Nbp2 was found together with Ptc1 and HOG pathway kinases. We have shown that Nbp2 is an adapter in this pathway, mediating interaction between Ptc1 and the Pbs2 MAP/ERK kinase in the HOG pathway.

Key Words

PP2C Ptc1 Nbp2 adapter MAPK MEK budding yeast