Defects in glycosylphosphatidylinositol (GPI) anchor synthesis activate Hog1 kinase and confer copper-resistance in Saccharomyces cerevisisae.
Las21/Gpi7 contains a heavy-metal-associated motif at its N-terminus. When this motif was disrupted by amino acid substitution, the cells acquired weak copper-resistance. We found that the previously isolated las21 mutants were strongly resistant to copper. Metallothionein is necessary for the expression of the copper-resistance of the las21 mutants. However, hyper-production of metallothionein is unlikely to be the cause of copper-resistance of the las21 mutants. Copper-sensitive mutants (collectively called Cus mutants) were isolated from the las21delta and characterized. One of the Cus genes was found to be PBS2, which encodes Hog1 MAP kinase kinase, indicating that the Hog1 MAP kinase pathway is needed for the expression of copper-resistance of the las21 mutants. As expected, the las21delta hog1delta strain was no longer copper-resistant. We found that Hog1 was constitutively activated in las21delta cells and in ssk1delta las21delta cells but not in sho1delta las21delta cells. Inactivation of either FSR2/MCD4 or MPC1/GPI13, both of which are involved in GPI anchor synthesis, like LAS21, caused a similar level of constitutive activation of Hog1 kinase and copper-resistance as found in the las21delta strain. The constitutive activation was canceled by introducing the sskl mutation, but not the sho1 mutation, in each GPI anchor mutant tested, suggesting that the defect in GPI anchor synthesis specifically affects the Slnl branch of the MAP kinase pathway. Since the wild-type cells grown in YPD containing 0.5 M NaCl do not show copper-resistance, mere activation of Hog1 is not sufficient for expression of copper-resistance. We propose that a defect in GPI anchor synthesis has multiple consequences, including activation of the Hog1 MAP kinase cascade and conferring copper-resistance.
Toh-e
A
A
Department of Biological Sciences, Graduate school of Science, The University of Tokyo, Hongo, Japan. toh-e@biol.s.u-tokyo.ac.jp
Oguchi
T
T
D002352:Carrier Proteins; D003300:Copper / Q000494:pharmacology*; D025141:Drug Resistance, Fungal / Q000235:genetics; D004789:Enzyme Activation; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis* / Q000235:genetics; D020935:MAP Kinase Signaling System; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D008668:Metallothionein / Q000235:genetics / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
C500769:CUP1-1 protein, S cerevisiae; D002352:Carrier Proteins; D017261:Glycosylphosphatidylinositols; C118052:MCD4 protein, S cerevisiae; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; C007121:copper thionein; D003300:Copper; D008668:Metallothionein; C413542:GPI13 protein, S cerevisiae; D017853:Phosphotransferases (Alcohol Group Acceptor); C088589:HOG1 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases