Dataset Information

SPATA2 links CYLD to the TNF-a receptor signaling complex and modulates the receptor signaling outcomes

ABSTRACT: TNF-a is a key regulator of innate immune and proinflammatory responses. However, the composition of the TNF-a receptor associated signaling complexes (TNF-RSC) and the architecture of the downstream signaling networks are incompletely understood. We employed quantitative mass spectrometry to demonstrate that TNF-a stimulation induces widespread protein phosphorylation and that the scope of phosphorylation expands in a temporal manner. TNF-a stimulation also induces rapid ubiquitylation of components of the TNF-RSC. Temporal analysis of the TNF-RSC composition identified SPATA2 as a novel component of the TNFRSC. The predicted PUB domain in the N-terminus of SPATA2 interacts with the USP domain of CYLD, whereas the C-terminus of SPATA2 interacts with HOIP. SPATA2 is required for recruitment of CYLD to the TNF-RSC. Downregulation of SPATA2 augments transcriptional activation of NF-jB and inhibits TNF-a-induced necroptosis, pointing to an important function of SPATA2 in modulating the outcomes of TNF-a signaling. Taken together, our study draws a detailed map of TNF-a signaling, identifies SPATA2 as a novel component of TNF-a signaling, and provides a rich resource for further functional investigations.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Petra Beli

LAB HEAD: Petra Beli

PROVIDER: PXD003627 | Pride | 2016-06-09

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20120828_QE3_LC1_SAW_SA_A549_TNFa_TiO2_1_10.raw	Raw
	20120828_QE3_LC1_SAW_SA_A549_TNFa_TiO2_1_11.raw	Raw
	20120828_QE3_LC1_SAW_SA_A549_TNFa_TiO2_1_12.raw	Raw
	20120828_QE3_LC1_SAW_SA_A549_TNFa_TiO2_1_7.raw	Raw
	20120828_QE3_LC1_SAW_SA_A549_TNFa_TiO2_1_8.raw	Raw

Items per page:

1 - 5 of 61

Similar Datasets

Project description:The bovine chromaffin cell (BCC) is a unique modelM-bM-^@M-^Ta highly homogeneous and accessible neuroendocrine cellM-bM-^@M-^Tin which to study gene regulation through first messenger-initiated signaling pathways that are specific to post-mitotic cells. BCCs were treated with tumor necrosis factor (TNF) or pituitary adenylate cyclase activating polypeptide (PACAP), two critical regulators of neural cell transcriptional programming during inflammation that act on TNFR2 and PAC1 receptors, respectively, in post-mitotic neuroendocrine cells. Transcripts which were significantly up regulated by either or both first messenger were identified from microarray analysis using two bovine oligonucleotide arrays (Affymetrix and Agilent) followed by statistical analysis with Partek Genomic suite. Microarray data were combined from the two arrays using qRT-PCR sampling validation, and the first-messenger transcriptome derived from TNF and PACAP signaling were compared. More than 90 percent of the genes up regulated either by TNF or PACAP were specific to a single first messenger. BioBase suite, DIRE and Opossum were used to identify common promoter/enhancer response elements that control the expression of TNF- or PACAP-stimulated genes. Bioinformatic analysis revealed that distinct groups of transcription factors control the expression of genes up regulated by either TNF or PACAP . Most of the genes up regulated by TNF contained response elements for members of the Rel transcription factor family, suggesting TNF-TNFR2 signaling mainly through the NF-kB signaling pathway. On the other hand, the PACAP regulated genes showed no enrichment for any single response element, containing instead response elements for combinations of transcription factors allowing activation through multiple signaling pathways, including cAMP, calcium and ERK, in neuroendocrine cells. Pharmacological strategies for mimicking neuroprotection by either PACAP or TNF in the context of CNS injury or degeneration in disease might focus on individual downstream gene activation pathways to achieve greater specificity in vivo. For our analysis we have used both Affymetrix and Agilent arrays to identify the genes that are regulated up or down by neuropeptide PACAP and TNF-alpha. On Affyemetrix platform we performed technical repeats with 3 arrays with untreated samples, 3 arrays with samples from PACAP treatment, and 3 array with samples from TNF-alpha treatment. On the Agilent platform we performed technical repeats with 3 arrays for each treatment hybridizing Cy3-labelled RNA from untreated samples and Cy5-labelled RNA from either TNF-alpha or PACAP treated samples.

Dataset Information

SPATA2 links CYLD to the TNF-a receptor signaling complex and modulates the receptor signaling outcomes

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets