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Competitive binding of STATs to receptor phospho-Tyr motifs accounts for altered cytokine responses in autoimmune disorders

ABSTRACT: In this study, we compared the effects of two cytokine treatments on the proteome of human Th-1 cells. We used saturating doses of murine single-chain IL-27 (EBI3+p28, 10nM) and HyperIL-6 (20nM) and continuously stimulated cells of three donors with the two cytokines for 24h or left untreated.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Leukocyte, Blood

SUBMITTER: Stephan Wilmes

LAB HEAD: Ignacio Moraga-Gonzalez

PROVIDER: PXD024188 | Pride | 2021-05-05

REPOSITORIES: Pride

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	SILAC_proteomics_24h_cytokine_stimulation_evaluated_metadata.xlsx	Xlsx
	SW451-1.raw	Raw
	SW451-2.raw	Raw
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	SW451-4.raw	Raw

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Publications

Competitive binding of STATs to receptor phospho-Tyr motifs accounts for altered cytokine responses.

Wilmes Stephan S Jeffrey Polly-Anne PA Martinez-Fabregas Jonathan J Hafer Maximillian M Fyfe Paul K PK Pohler Elizabeth E Gaggero Silvia S López-García Martín M Lythe Grant G Taylor Charles C Guerrier Thomas T Launay David D Mitra Suman S Piehler Jacob J Molina-París Carmen C Moraga Ignacio I

eLife 20210419

Cytokines elicit pleiotropic and non-redundant activities despite strong overlap in their usage of receptors, JAKs and STATs molecules. We use IL-6 and IL-27 to ask how two cytokines activating the same signaling pathway have different biological roles. We found that IL-27 induces more sustained STAT1 phosphorylation than IL-6, with the two cytokines inducing comparable levels of STAT3 phosphorylation. Mathematical and statistical modeling of IL-6 and IL-27 signaling identified STAT3 binding to ...[more]

PMID: 33871355

Similar Datasets

Competitive binding of STATs to receptor phospho-Tyr motifs accounts for altered cytokine responses in autoimmune disorders

Project description:In this study, we compared the effects of two cytokine treatments on the proteome of human Th-1 cells. We used saturating doses of murine single-chain IL-27 (EBI3+p28, 10nM) and HyperIL-6 (20nM) and continuously stimulated cells of three donors with the two cytokines for 15min or left untreated.

2021-05-05 | PXD024657 | Pride

IL6-regulated CDk8-mediated phosphoproteome of human primary Th1 cells

Project description:Cytokines are highly pleiotropic ligands that critically contribute to a balanced immune response. We have an incomplete understanding of how cytokines elicit their functional pleiotropy, which has limited their therapeutic efficacy. Here, using Interleukin-6 (IL-6) as a model system, we have performed detailed phosphoproteomic and transcriptomic studies in human CD4 + T helper (Th)-1 cells to address the molecular bases defining cytokine functional pleiotropy. We have identified cyclin dependent kinase (CDK)8 as a new negative regulator of transcriptional activities of signal transducer and activator of transcription (STAT)3. We found that CDK8 is a major regulator of the IL-6 phosphoproteome and interacts with STAT3 in the nucleus upon IL-6 stimulation. Inhibition of CDK8 activity, using specific small molecules inhibitors, reduced the IL-6-induced phosphoproteome by 23% in Th-1 cells, including STAT3 S727 phosphorylation. STAT3 binding to target DNA sites in the genome was increased upon CDK8 inhibition, which resulted in a concomitant increase in STAT3 mediated transcriptional activity. Importantly, inhibition of CDK8 activity under Th-17 polarizing conditions resulted in an enhancement of Th-17 differentiation. Our results support a model where CDK8 regulates STAT3 transcriptional processivity via modulation of its gene loci resident time, critically contributing to diversification of IL-6 responses.

2020-12-23 | PXD020964 | Pride

Analysis of HeLa cells after transfection with miR-1, miR-124 and miR-181.

Project description:Analysis of HeLa cells at 24 hours after transfection with wild type miR-1, miR-124, miR-181 versus control transfected HeLa cells. Results were compared to protein down-regulation at 48 hours measured by SILAC-MS. Analysis of HeLa cells at 24 hours after transfection with wild type miR-1, miR-124, miR-181 versus control transfected HeLa cells. Results were compared to protein down-regulation at 48 hours measured by SILAC-MS.

2010-05-25 | E-GEOD-11968 | biostudies-arrayexpress

Analysis of small RNAs in murine neutrophils by Solexa/Illumina genome analyzer

Project description:To examine the effects of microRNAs including miR-223 on murine neutrophil function, small RNAs cloning and sequencing from neutrophils were performed. Neutrophils were isolated from a wild-type mouse and total RNAs were prepared. Small RNAs were cloned and sequenced by Solexa/Illumina genome analyzer.

2008-07-31 | E-GEOD-12074 | biostudies-arrayexpress

HeLa Adenovirus timecourse

Project description:A timecourse of adenovirus infection of human HeLa cells, using RNA-seq

2012-10-27 | E-MTAB-1277 | biostudies-arrayexpress

Transcription profiling of human hematopoietic stem cells

Project description:Successful ex vivo expansion of hematopoietic stem cells (HSCs) would greatly benefit the treatment of disease and the understanding of critical questions of stem cell biology. Microarray studies showed that the HSC supportive mouse fetal liver CD3+ cells specifically expressed angiopoietin-like 2 (Angptl2) and angiopoietin-like 3 (Angptl3). When highly enriched HSCs were cultured in the presence of Angptl2 or Angptl3 together with saturating levels of other growth factors for 10 days, a 24 or 30 fold net expansion of long-term HSCs was observed by reconstitution analysis. The coiled-coil domain of Angptl2 mediated the effects of Angptl2 on HSC expansion. Furthermore, angiopoietin-like 5, angiopoietin-like 7, and microfibril-associated glycoprotein 4 also supported expansion of HSCs in culture. Experiment Overall Design: Cultured 3 different cell types Experiment Overall Design: -> Total RNA isolation -> dscDNA -> Biotinylated cRNA -> hybridization to Affymetrix U74Bv2 and U74Cv2 mouse chips

2008-06-12 | E-GEOD-3609 | biostudies-arrayexpress

Sinorhizobium meliloti Ser/Thr/Tyr phosphoproteome

Project description:Sinorhizobium meliloti, a facultative microsymbiont of alfalfa, should fine-tune its cellular processes to live saprophytically in soils characterized with limited nutrients and diverse stresses. In this study, TiO2 enrichment and LC-MS/MS were used to uncover the site-specific Ser/Thr/Tyr phosphoproteome of S. meliloti in minimum medium at stationary phase. There are a total of 96 unique phosphorylated sites, with a Ser/Thr/Tyr distribution of 65:32:5, in 78 proteins. Phosphoproteins identified in S. meliloti showed a wide distribution pattern regarding to functional categories, such as replication, transcription, translation, posttranslational modification, transport and metabolism of amino acids, carbohydrate, inorganic ion, succinoglycan etc. Ser/Thr/Tyr phosphosites identified within the conserved motif in proteins of key cellular function indicate a crucial role of phosphorylation in modulating cellular physiology. Moreover, phosphorylation events potentially involved in rhizobial adaptation to diverse stresses were also discussed, such as those identified in SMa0114 and PhaP2 (polyhydroxybutyrate synthesis), ActR (pH stress and microaerobic adaption), SupA (potassium stress), chaperonin GroEL2 (viability and potentially symbiosis), and ExoP (succinoglycan synthesis and secretion). These Ser/Thr/Tyr phosphosites identified herein would be helpful for our further investigation and understanding of the role of phosphorylation in rhizobial physiology.

2015-10-05 | PXD002064 | Pride

Transcriptome of melanoma cells from two mouse models, Tyr:NRasQ61K and Tyr:Rack1-HA, Tyr:NRasQ61K.

Project description:The transcriptome sequencing of melanoma cells from two mouse models differing in the expression level of the scaffold protein Receptor for activated C kinase (RACK1) are presented. Primary melanoma cells were harvested from Tyr:NRasQ61K; Pax3GFP/+ mice, with or without the Tyr:Rack1-HA transgene. Cells were cultured and infected with scramble shRNA or Rack1-targeting shRNA, on technical triplicates of viral infection. Libraries were prepared by selecting polyadenylated mRNAs and RNA Sequencing (RNASeq) was performed. Samples are described in the SRA portal (SRP096162) and FASTQ files have been deposited in Sequence Read Archive (accession numbers: SRR5150106 to SRR5150117). The interpretation of these data is presented in the following research article: "RACK1 cooperates with NRASQ61K to promote melanoma in vivo" (Campagne et al., 2017, doi: 10.1016/j.cellsig.2017.03.015) [1].

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Detection of CHMP2B methylation

Project description:Characterization of CHMP2B methylation by the lysine methyl transferase SMYD2 : identification of the lysine methylated by SMYD2

2024-03-18 | PXD041760 | Pride

Mouse tissue-resident macrophage expression profiles

Project description:We show that macrophages from murine spleen, liver and peritoneum display dramatically different expression profiles. Clusters of genes were found to represent unique biological functions related to adhesion, antigen presentation, phagocytosis, lipid metabolism and signal transduction. Some gene families, such as integrins, are differentially expressed among the macrophages resident in different tissues, suggesting that the tissue of residence influences their biological function. 18 FACS sorted mouse tissue-resident macrophge samples. 6 cell types, 3 biological replicates per cell type.

2010-09-22 | E-GEOD-24258 | biostudies-arrayexpress