Dataset Information

Identifying protein kinase specific effectors of the osmostress response in yeast_part2

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: PXD004295 | Pride | 2017-02-01

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20130607_Velos2_RSLC5_Reiter_Ammerer_MFPL_insol_hog1_silac_0stress_elu1col1_1per_tio2-01.msf	Msf
	20130607_Velos2_RSLC5_Reiter_Ammerer_MFPL_insol_hog1_silac_0stress_elu1col1_1per_tio2.raw	Raw
	20130607_Velos2_RSLC5_Reiter_Ammerer_MFPL_insol_hog1_silac_0stress_elu1col2_1per_tio2-01.msf	Msf
	20130607_Velos2_RSLC5_Reiter_Ammerer_MFPL_insol_hog1_silac_0stress_elu1col2_1per_tio2.raw	Raw
	20130613_Velos2_RSLC5_Reiter_Ammerer_MFPL_insol_SCX2_hog1_silac_inh_0minstress_5per.msf	Msf

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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]

PMID: 28270554

Similar Datasets

Project description:Schaber2012 - Hog pathway in yeast The high osmolarity glycerol (HOG) pathway in the yeast Saccharomyces cerevisiae is one of the best-studied mitogen-activated protein kinase (MAPK) pathways and serves as a prototype signalling system for eukaryotes. This pathway is necessary and sufficient to adapt to high external osmolarity. A key component of this pathway is the stress-activated protein kinase (SAPK) Hog1, which is rapidly phosphorylated by the SAPK kinase Pbs2 upon hyper-osmotic shock, and which is the terminal kinase of two parallel signalling pathways, subsequently called the Sho1 branch and the Sln1 branch, respectively. Ensemble modelling (192 models) is used to study the yeast HOG pathway, a prototype for eukaryotic mitogen-activated kinase signalling systems. The best fit model (Model Nr.22: described here) provides new insights into the function of this system, some of which are then experimentally validated. This model is described in the article: Modelling reveals novel roles of two parallel signalling pathways and homeostatic feedbacks in yeast. Schaber J, Baltanas R, Bush A, Klipp E, Colman-Lerner A. Mol Syst Biol. 2012 Nov 13;8:622. Abstract: The high osmolarity glycerol (HOG) pathway in yeast serves as a prototype signalling system for eukaryotes. We used an unprecedented amount of data to parameterise 192 models capturing different hypotheses about molecular mechanisms underlying osmo-adaptation and selected a best approximating model. This model implied novel mechanisms regulating osmo-adaptation in yeast. The model suggested that (i) the main mechanism for osmo-adaptation is a fast and transient non-transcriptional Hog1-mediated activation of glycerol production, (ii) the transcriptional response serves to maintain an increased steady-state glycerol production with low steady-state Hog1 activity, and (iii) fast negative feedbacks of activated Hog1 on upstream signalling branches serves to stabilise adaptation response. The best approximating model also indicated that homoeostatic adaptive systems with two parallel redundant signalling branches show a more robust and faster response than single-branch systems. We corroborated this notion to a large extent by dedicated measurements of volume recovery in single cells. Our study also demonstrates that systematically testing a model ensemble against data has the potential to achieve a better and unbiased understanding of molecular mechanisms. This model is hosted on BioModels Database and identified by: MODEL1209110001 . To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models . To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.