Proteomics

Dataset Information

SILAC Mass Spectrometry of U2AF2 Interactome in Resting and Activated CD4 T cells


ABSTRACT: T cell activation leads to dramatic changes in cellular phenotype. We used activated human CD4 T cells to study how RNA binding proteins define the post-transcriptional landscape. Using RIPseq, we identified the RNA interactome of U2AF2 and show that U2AF2 binds the majority of transcripts that are differentially expressed and/or alternatively spliced during activation. Using RIP mass spectrometry, a unique protein interactome centered on U2AF2 is assembled by activation and comprised of both directly bound central members (RNAse-resistant) and indirectly bound peripheral members (RNAse-sensitive). Knocking down specific U2AF2 interactome members (U2AF1, SYNCRIP, SRRM2, ILF2) selectively affects cytokine secretion and activation markers. The expression and/or alternative splicing of transcripts important for immune cell function are also affected by knocking down these interactome members, both peripheral and central. Furthermore, we show that knockdown of interactome members can affect the proteins and transcripts bound to U2AF2, altering the transcriptome of activated T cells. Our work highlights the importance of understanding the assembly of RNA binding protein complexes as regulators of T cell activation and function.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell, Cell Culture

SUBMITTER: Thomas Whisenant  

LAB HEAD: Daniel R. Salomon, M.D.

PROVIDER: PXD001846 | Pride | 2016-04-27

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
U2AF2IP_SILAC.xml Xml
U2IP_RestAct_SILAC_1.mzml Mzml
U2IP_RestAct_SILAC_2.mzml Mzml
U2IP_RestAct_SILAC_3.mzml Mzml
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