Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

In depth characterization of the S. aureus phosphoproteome reveals new targets of Stk1

ABSTRACT: Staphylococcus aureus is the leading cause of infections worldwide and infection results in a variety of diseases. As of no surprise, phosphorylation is a major game player in signaling cascades and has been shown to be involved in S. aureus virulence. Albeit long neglected, eukaryotic-like serine/threonine kinases have been implicated in these complex signaling cascades. Due to the sub-stoichiometric nature of protein phosphorylation and a lack of suitable analysis tools, the knowledge of these cascades is however, to date, still limited. Here, were apply an optimized protocol for efficient phosphopeptide enrichment via Fe3+-IMAC followed by LC-MS/MS to get a better understanding of the impact of protein phosphorylation on the complex signaling networks involved in pathogenicity. By profiling a serine/threonine kinase and phosphatase mutant from a methicillin-resistant S. aureus mutant library, we generated the most comprehensive phosphoproteome dataset of S. aureus to date, aiding a better understanding of signaling in bacteria.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Staphylococcus Aureus (strain Usa300) Bacillus Subtilis Subsp. Subtilis Str. 168

SUBMITTER: Nadine Prust

LAB HEAD: Simone Lemeer

PROVIDER: PXD020226 | Pride | 2020-12-21

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Extensive arginine phosphorylation in Staphylococcus aureus is influences by Stp1

Project description:Ever since the identification of the first protein-arginine kinase, McsB, in B. subtilis arginine phosphorylation gained more attention. However, the analysis of phosphorylations especially phosphormaidates comes along with several challenges. The sub-stoichiometric nature of protein phosphorylation requires proper enrichment strategies prior to LC-MS/MS analysis and the acid instability of phosphoramidates was long though to impede those enrichments. Further good spectral quality is required which can be reduced by the presence of neutral losses of phosphoric acid upon higher energy collision induced dissociation. Adaptation of common enrichment strategies to less acidic conditions and the use of electron-transfer dissociation allowed the identification of more than 200 pArg sites in B. subtilis and S. aureus so far. Here we show that pArg is stable enough for commonly used Fe3+-IMAC enrichment followed by LC-MS/MS and that HCD is still the gold standard for phosphoproteomics. By profiling a serine/ threonine kinase (Stk1) and phosphatase (Stp1) mutant from a methicillin-resistant S. aureus mutant library, we identified 1062 pArg sites and thus the most comprehensive arginine phosphoproteome to date. Using synthetic phosphoarginine peptides we validated the presence and localization of arginine phosphorylation in S. aureus and lastly, we could show that Stp1 is influencing the arginine phosphoproteom without being a protein-arginine phosphatase.

2022-06-09 | PXD026981 | Pride

Expression data from P14 TCR cytotoxic T cells overexpressing HDAC7 phosphorylation deficient mutant

Project description:The present study reports an unbiased analysis of the cytotoxic T cell serine-threonine phosphoproteome using high resolution mass spectrometry. Approximately 2,000 phosphorylations were identified in CTLs of which approximately 450 were controlled by TCR signaling. A significantly overrepresented group of molecules identified in the phosphoproteomic screen were transcription activators, co-repressors and chromatin regulators. A focus on the chromatin regulators revealed that CTLs have high expression of the histone deacetylase HDAC7 but continually phosphorylate and export this transcriptional repressor from the nucleus. HDAC7 dephosphorylation results in its nuclear accumulation and suppressed expression of genes encoding key cytokines, cytokine receptors and adhesion molecules that determine CTL function. The screening of the CTL phosphoproteome thus reveals intrinsic pathways of serine-threonine phosphorylation that target chromatin regulators in CTLs and determine the CTL functional program. We used Affymetrix microarray analysis to explore the molecular basis for the role of HDAC7 in CTLs and the impact of GFP-HDAC7 phosphorylation deficient mutant expression on the CTL transcriptional profile. In vitro generated P14 TCR cytotoxic T cells were retrovirally infected with a construct encoding GFP-HDAC7 phosphorylation deficient mutant, sorted in base of GFP expression (GFP positive and GFP negative) and processed for microarray analysis in three biological replicas.

2011-03-13 | E-GEOD-27092 | biostudies-arrayexpress

R406-Inhibition of Syk-Cn-NFAT Signaling Axis Promotes Chemical Reprogramming via Metabolic Rewiring and H2S Production

Project description:Small molecules not only are convenient and useful for controlling cell fates, but also can provide better understanding of the molecular mechanisms of cell-fate transitions. Here, we identified that spleen tyrosine kinase (Syk) inhibitor R406 could significantly promote the early stage of mouse chemical reprogramming. Furthermore, R406 alleviated the Syk-calcineurin (Cn)-nuclear factor of activated T cells (NFAT) signaling cascade-mediated suppression of glycine, serine and threonine metabolic genes transcriptionally, which is previously unrecognized. In turn, R406 upregulated metabolites of glycine, serine and threonine metabolism and downstream transsulfuration cysteine biosynthesis pathway and cysteine metabolism. Subsequently, increased cellular hydrogen sulfide (H2S) after R406 treatment downregulated oxidative phosphorylation (OXPHOS) and reactive oxygen species (ROS), modulated redox homeostasis, and significantly enhanced chemical reprogramming. In sum, our studies have not only improved the chemical reprogramming technique, but also identified interesting molecular mechanisms of chemical-induced pluripotent reprogramming, which hold great potentials in regenerative medicine.

2021-03-14 | GSE154687 | GEO

PrrA modulation of Mycobacterium tuberculosis response to nitric oxide is critically regulated by serine/threonine protein kinases

Project description:The purpose of this study was to understand how prevention of serine/threonine protein kinase (STPK) phosphorylation of PrrA impacts PrrA modulation of M. tuberculosis transcriptional response to nitric oxide.

2022-06-23 | GSE198999 | GEO

PrrA modulation of Mycobacterium tuberculosis response to acidic pH and high chloride levels is critically regulated by serine/threonine protein kinases

2022-06-23 | GSE199000 | GEO

Gene expression profiles in GSK-3beta Ser9Ala knock-in mouse heart

Project description:GSK-3 is a serine-threonine protein kinase and consists of two isoforms, GSK-3alpha and GSK-3beta. GSK-3alpha and GSK-3beta are inactivated by phosphorylation at Ser21 and Ser9, respectively. GSK-3alpha Ser21Ala and GSK-3beta Ser9Ala phosphorylation-resistant knock-in mice have constitutively active form of GSK-3.

2019-03-11 | GSE112159 | GEO

Gene expression profiles in GSK-3alpha Ser21Ala knock-in mouse heart

2019-03-11 | GSE112158 | GEO

Transcriptome analysis of the eukaryotic-type ser/thr kinase PknB in Staphylococcus aureus

Project description:The function of the Staphylococcus aureus eukaryotic-like serine/threonine protein kinase PknB was investigated by transcriptome analysis using DNA-microarray technology and biochemical assays. The transcriptional profile reveals a strong regulatory impact of PknB on the expression of genes encoding proteins which are involved in purine and pyrimidine biosynthesis, cell wall metabolism, autolysis, and glutamine synthesis.

2010-03-11 | GSE15346 | GEO

Expression data from P14 TCR cytotoxic T cells overexpressing HDAC7 phosphorylation deficient mutant

2011-03-13 | GSE27092 | GEO

IRS1 phosphorylation underlies the non-stochastic probability of cancer cells to persist during EGFR inhibition therapy [Exp3]

Project description:We found that differential serine/threonine phosphorylation of the insulin receptor substrate 1 (IRS1) protein determines the chance to persist (CTP) of lung and of head and neck cancer cells under EGFR inhibition, both in vitro and in vivo.

2021-08-09 | GSE178977 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data