Genetics, Vol. 159, 1479-1489, December 2001, Copyright © 2001

Sit4p Protein Phosphatase Is Required for Sensitivity of Saccharomyces cerevisiae to Kluyveromyces lactis Zymocin

Daniel Jablonowskia, Andrew R. Butlerb, Lars Fichtnera, Donald Gardinerb, Raffael Schaffratha, and Michael J. R. Starkb
a Institut für Genetik, Martin-Luther Universität Halle-Wittenberg, D-06120 Halle (Salle), Germany
b Division of Gene Regulation & Expression, School of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom

Corresponding author: Michael J. R. Stark, Division of Gene Regulation & Expression, School of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom., m.j.r.stark{at}dundee.ac.uk (E-mail)

Communicating editor: P. RUSSELL

We have identified two Saccharomyces cerevisiae genes that, in high copy, confer resistance to Kluyveromyces lactis zymocin, an inhibitor that blocks cells in the G1 phase of the cell cycle prior to budding and DNA replication. One gene (GRX3) encodes a glutaredoxin and is likely to act at the level of zymocin entry into sensitive cells, while the other encodes Sap155p, one of a family of four related proteins that function positively and interdependently with the Sit4p protein phosphatase. Increased SAP155 dosage protects cells by influencing the sensitivity of the intracellular target and is unique among the four SAP genes in conferring zymocin resistance in high copy, but is antagonized by high-copy SAP185 or SAP190. Since cells lacking SIT4 or deleted for both SAP185 and SAP190 are also zymocin resistant, our data support a model whereby high-copy SAP155 promotes resistance by competition with the endogenous levels of SAP185 and SAP190 expression. Zymocin sensitivity therefore requires a Sap185p/Sap190p-dependent function of Sit4p protein phosphatase. Mutations affecting the RNA polymerase II Elongator complex also confer K. lactis zymocin resistance. Since sit4{Delta} and SAP-deficient strains share in common several other phenotypes associated with Elongator mutants, Elongator function may be a Sit4p-dependent process.




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