Premium
Functional and genomic analyses of blocked protein O‐mannosylation in baker's yeast
Author(s) -
Arroyo Javier,
Hutzler Johannes,
Bermejo Clara,
Ragni Enrico,
GarcíaCantalejo Jesús,
Botías Pedro,
Piberger Heidi,
Schott Andrea,
Sanz Ana Belén,
Strahl Sabine
Publication year - 2011
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2011.07537.x
Subject(s) - biology , endoplasmic reticulum associated protein degradation , saccharomyces cerevisiae , mutant , unfolded protein response , yeast , gene , microbiology and biotechnology , genetics
Summary O‐mannosylation is a crucial protein modification in eukaryotes that is initiated by the essential family of protein O‐mannosyltransferases (PMTs). Here we demonstrate that in the model yeast Saccharomyces cerevisiae rhodanine‐3‐acetic acid derivatives affect members of all PMT subfamilies. Specifically, we used OGT2468 to analyse genome‐wide transcriptional changes in response to general inhibition of O‐mannosylation in baker's yeast. PMT inhibition results in the activation of the cell wall integrity (CWI) pathway. Coinciding, the mitogen‐activated kinase Slt2p is activated in vivo and CWI pathway mutants are hypersensitive towards OGT2468. Further, induction of many target genes of the unfolded protein response (UPR) and ER‐associated protein degradation (ERAD) is observed. The interdependence of O‐mannosylation and UPR/ERAD is confirmed by genetic interactions between HAC1 and PMT s, and increased degradation of the ERAD substrate Pdr5p* in pmt Δ mutants. Transcriptome analyses further suggested that mating and filamentous growth are repressed upon PMT inhibition. Accordingly, in vivo mating efficiency and invasive growth are considerably decreased upon OGT2468 treatment. Quantitative PCR and ChIP analyses suggest that downregulation of mating genes is dependent on the transcription factor Ste12p. Finally, inhibitor studies identified a role of the Ste12p‐dependent vegetative signalling cascade in the adaptive response to inhibition of O‐mannosylation.