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Nigericin-Triggered Phosphodynamics in Inflammasome Formation and Pyroptosis

ABSTRACT: Innate immune signaling relies on phosphorylation cascades for effective responses. While TLR4 stimulation has been well-studied, research on signaling after inflammasome triggers like nigericin is limited. We performed time-resolved phosphoproteomic profiling to explore kinase signaling downstream of nigericin and discovered rapid changes in phosphorylation, particularly involving immune and MAPK pathways. Notably, we identified fluxes in PKC signaling and calcium-responsive proteins, along with phospho-modified metabolic pathways, indicating a role in immunometabolic regulation. Additionally, significant changes in DNA reorganization and damage response proteins linked to pyknosis were observed. Temporal clustering of phospho-dynamics revealed new shifts in signaling cascades after nigericin treatment, underscoring changes in cellular behavior related to inflammatory events

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Macrophage

DISEASE(S): Disease Free

SUBMITTER: Aleksandra Nita-Lazar

LAB HEAD: Aleksandra Nita-Lazar

PROVIDER: PXD062818 | Pride | 2025-08-27

REPOSITORIES: Pride

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			Action	DRS
	GLOBAL_PROTEOME_DISCOVERER_ANALYSIS.msf	Msf
	Global_Analysis_InputFiles.txt	Txt
	Global_Analysis_MSMSSpectrumInfo.txt	Txt
	Global_Analysis_PSMs.txt	Txt
	Global_Analysis_PeptideGroups.txt	Txt

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Project description:Human histomonocytic U937 cells exhibit macrophage-like properties after phorbol ester stimulation. Accumulating evidence demonstrated that remarkable number of transcripts changed in their amount at total RNA levels. However, post-transcriptional regulation has not been fully elucidated in this model. Thus, we compared expression profiles between polysomal and cytoplasmic RNAs before and after phorbol 12-myristate 13-acetate stimulation (48h, 32nM). Table 1: Detailed description of the expression data of human histomonocytic cell line U937 cells. Data of total cytoplasmic and polysomal fractions before and after PMA stimulation is shown. This table contains all spot data except: 1) saturated spots, 2) undetectable spots, 3) stained spots, and 4) spots only detected in less than one samples. Column A: Gene symbol (âI_xxxxxxâ is corresponding to EST clones); Column B: Genbank accession no. Column C: Description; Column D: Average value of expression levels in total cytoplasmic fraction in the absence of PMA; Column E: Average value of expression levels in total cytoplasmic fraction in the presence of PMA (32nM, 48h); Column F: Average value of expression levels in polysomal fraction in the absence of PMA; Column G: Average value of expression levels in polysomal fraction in the presence of PMA; Column H: Expression ratio of polysome, PMA (-) to cytoplasm, PMA(-); Column I: Expression ratio of polysome, PMA (+) to cytoplasm, PMA(+); Column J: Expression ratio of cytoplasm, PMA (+) to cytoplasm, PMA(-); Column K: Expression ratio of polysome, PMA (+) to polysome, PMA(-); Column L: Different expression between polysome, PMA (-) and cytoplasm, PMA(-): different=1; Column M: Different expression between polysome, PMA (+) and cytoplasm, PMA(+): different=1; Column N: Different expression between cytoplasm, PMA (+) and cytoplasm, PMA(-): different=1; Column O: Different expression between polysome, PMA (+) and polysome, PMA(-): different=1; ; Table 2: Candidates post-transcriptionally regulated by PMA. We extracted 105 transcripts whose expression levels were altered only in either fraction accompanied by changes in polysome/cytoplasm expression ratio. Column A: Gene symbol (âI_xxxxxxâ is corresponding to EST clones); Column B: Genbank accession no. Column C: Description; Column D: Average value of expression levels in total cytoplasmic fraction in the absence of PMA; Column E Average value of expression levels in total cytoplasmic fraction in the presence of PMA; Column F: Average value of expression levels in polysomal fraction in the absence of PMA; Column G: Average value of expression levels in polysomal fraction in the presence of PMA; Column H: Expression ratio of polysome, PMA (-) to cytoplasm, PMA(-); Column I: Expression ratio of polysome, PMA (+) to cytoplasm, PMA(+); Column J: Expression ratio of cytoplasm, PMA (+) to cytoplasm, PMA(-); Column K: Expression ratio of polysome, PMA (+) to polysome, PMA(-)

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Nigericin-Triggered Phosphodynamics in Inflammasome Formation and Pyroptosis

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