Proteomics

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Identifying protein kinase specific effectors of the osmostress response in yeast_part4


ABSTRACT: Saccharomyces cerevisiae reacts to elevated external osmolarity by several cellular responses. Thereby, the main adaptive signaling activity relies on the high-osmolarity glycerol (HOG) pathway, whose core architecture is closely related to the mammalian p38 MAPK pathway. To identify novel target proteins of its MAP kinase Hog1, we applied an MS-based high throughput analysis measuring the impact of Hog1 activation as well as inhibition on the budding yeast phosphoproteome. In addition, we analyzed how a deletion of RCK2, a known effector protein kinase target of Hog1, modulates the normal stress induced phosphorylation pattern. Our results provide an overview on the diversity of cellular functions that are directly and indirectly affected by the activity of the pathway and allow a clear assessment of Hog1 independent events affected by osmotic stress. Analyzing the modulation of S/T-P motifs, we could extend the number of Hog1 putative direct targets that were subsequently validated by an in vivo interaction assay. It also appears that Rck2, a downstream kinase of Hog1, acts as a central hub for many Hog1-mediated secondary phosphorylation events.

INSTRUMENT(S): LTQ Orbitrap Velos, Q Exactive

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Wolfgang Reiter  

LAB HEAD: Gustav Ammerer

PROVIDER: PXD004297 | Pride | 2017-02-01

REPOSITORIES: Pride

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Publications

Identifying protein kinase-specific effectors of the osmostress response in yeast.

Romanov Natalie N   Hollenstein David Maria DM   Janschitz Marion M   Ammerer Gustav G   Anrather Dorothea D   Reiter Wolfgang W  

Science signaling 20170307 469


The budding yeast <i>Saccharomyces cerevisiae</i> reacts to increased external osmolarity by modifying many cellular processes. Adaptive signaling relies primarily on the high-osmolarity glycerol (HOG) pathway, which is closely related to the mammalian p38 mitogen-activated protein kinase (MAPK) pathway in core architecture. To identify target proteins of the MAPK Hog1, we designed a mass spectrometry-based high-throughput experiment to measure the impact of Hog1 activation or inhibition on the  ...[more]

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