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Regulation of Far11 phosphorylation by Ppg1 phosphatase

ABSTRACT: Ppg1 phosphatase post-translationally regulates Far11. To identify Far11 phosphorylation sites regulated by Ppg1, we immunopurified Far11 from wild-type and Ppg1 catalytically inactive (Ppg1-H111N) cells. We used this immunopurified Far11 for proteomics and identified Ppg1 dependent Far11 phosphorylation sites.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Shreyas Niphadkar

LAB HEAD: Sunil Laxman

PROVIDER: PXD049989 | Pride | 2024-02-29

REPOSITORIES: Pride

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Project description:Differentiation of CD4+T-cells into effector subsets is a critical component of the adaptive immune system and an incorrect response can lead to autoimmunity or immune deficiency. Cellular differentiation including T-cell differentiation is accompanied by large-scale epigenetic remodeling, including changes in DNA methylation at key regulators of T-cell differentiation. The TET family of enzymes were recently shown to be able to catalyse methylated cytosine (5mC) into 5-hydroxymethylcytosine (5hmC) enabling a pathway of active removal of DNA methylation. Here, we characterize 5hmC, 5mC and transcriptional dynamics during human CD4+T-cell polarisation in a time series approach and relate these changes to profiles in ex-vivo CD4+memory subsets. We observed large-scale remodelling during early CD4+T-cell differentiation which was predictive of subsequent changes during late time points, these changes were also related to disease associated regions which we show can act as functional regulatory elements. This dataset was designed to assess how TET1 affects gene expression during human CD4+T-cell differentiation. We observed that TET1 was highly expressed in primary lymphocytes compared to other human tissues and loss of TET1 gene expression was seen within 6 hours of naÃ¯ve CD4+T-cell activation. Primary human naÃ¯ve T-cells were transfected with different TET1 containing plasmids and polarized towards Th1 for 24h. When comparing cells transfected with full length TET1 to catalytically inactive TET1, we observed mis-expression of several cytokine/chemokine genes related to T-cell differentiation. This was not observed for cells expressing the catalytic domain of TET1. Note that the expression array only targets the DNA binding domain of the TET1 gene, therefore we validated ectopic expression of TET1 by qPCR targeting the catalytic domain. This submission contains data from Transcription profiling by array of primary human CD4+T-cells overexpressing TET1 polarized towards Th1. See related experiments: E-MTAB-4686, E-MTAB-4687, E-MTAB-4688, E-MTAB-4689.

			Action	DRS
	Ppg1_H111N_1.raw	Raw
	Ppg1_H111N_1.xml	Xml
	Ppg1_H111N_2.raw	Raw
	Ppg1_H111N_2.xml	Xml
	Ppg1_H111N_3.raw	Raw

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Regulation of Far11 phosphorylation by Ppg1 phosphatase

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