Proteomics

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Inverted chromatin immunoprecipitation identifies Tcf1 as a negative regulator of Foxp3 expression


ABSTRACT: Regulatory T (Treg) cells are important regulators of the immune system and have versatile functions for the homeostasis and repair of tissues. They express the forkhead box transcription factor Foxp3 as a lineage-defining protein. In mature Treg cells, the Foxp3 core promoter is unmethylated indicating that this area could harbor a transcription factor complex to initiate or repress gene expression, respectively. We used an unbiased method to identify Foxp3-promoter-binding transcription factors (TFs) by inverted chromatin immunoprecipitation (IP) followed by quantitative mass spectrometry. We identified several candidate factors which showed Foxp3-promoter suppressive capacity, one of which was T-cell factor 1 (Tcf1). Using viral overexpression and CRISPR/Cas knockout studies, we identified Tcf1 as a repressor of Foxp3 expression in primary conventional CD4 T cells (Tconv). In Tcf1-deficient animals, increased levels of Foxp3intermediateCD25negative T cells were identified in secondary lymphoid tissues, implicating that Tcf1 protects Foxp3-negative T cells from inadvertent Foxp3 expression.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): T Cell

SUBMITTER: Mahmoudreza Rafiee  

LAB HEAD: Jeroen Krijgsveld

PROVIDER: PXD018764 | Pride | 2020-05-26

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
121017_KE_L1_M1_10_5.raw Raw
121017_KE_L1_M1_11_5.raw Raw
121017_KE_L1_M1_12_5.raw Raw
121017_KE_L1_M1_1_5.raw Raw
121017_KE_L1_M1_2_5.raw Raw
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Publications


Regulatory T cells are important regulators of the immune system and have versatile functions for the homeostasis and repair of tissues. They express the forkhead box transcription factor Foxp3 as a lineage-defining protein. Negative regulators of Foxp3 expression are not well understood. Here, we generated double-stranded DNA probes complementary to the Foxp3 promoter sequence and performed a pull-down with nuclear protein in vitro, followed by elution of bound proteins and quantitative mass sp  ...[more]

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