Dataset Information

Intestinal Paneth cell differentiation relies on asymmetric regulation of Wnt signaling by Daam1/2

ABSTRACT: Purification of RNF43 interactors was performed with the following 4 different constructs: (i) empty vector (EV) as a control for background contaminants/non-specific protein binding; (ii) full-length RNF43 protein (RNF43 FL); (iii) deleted version of RNF43 harboring only the PA-TA domain (RNF43 Δ1); and (iv) deleted version of RNF43 harboring only the PA-TA-RING domain (RNF43 Δ3). HEK293T cells were transfected with pcDNA3-FLAG-HA- RNF43 (FL, Δ1 or Δ3).

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Bon-Kyoung Koo

PROVIDER: PXD043151 | JPOST Repository | Thu Oct 19 00:00:00 BST 2023

REPOSITORIES: jPOST

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Source:

			Action	DRS
	20110312_RNF43_IP_All.sf3	Other
	OR1_110312_TY_RNF43_CTRL01.RAW	Raw
	OR1_110312_TY_RNF43_CTRL01.mgf	Mgf
	OR1_110312_TY_RNF43_CTRL02.RAW	Raw
	OR1_110312_TY_RNF43_CTRL02.mgf	Mgf

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Project description:We have previously shown that adenoviral expression of peroxisome proliferator-activated receptors (PPARs) leads to rapid establishment of transcriptionally active complexes and activation of target gene expression within 5-8 h following transduction. Here we have used the adenoviral delivery system combined with expression array analysis to identify novel putative PPARgamma target genes in murine fibroblasts and to determine the role of the A/B-domain in PPARgamma mediated transactivation of genomic target genes. Of the 257 genes found to be induced by PPARgamma2 expression, only 25 displayed A/B-domain dependency, i.e. significantly reduced induction in the cells expressing the truncated PPARgamma lacking the A/B-domain (PPARgammaCDE). Nine of the 25 A/B-domain dependent genes were involved in lipid storage and in line with this, triglyceride accumulation was considerably decreased in the cells expressing PPARgammaCDE compared to cells expressing full length PPARgamma2. Using chromatin immunoprecipitation (ChIP) we demonstrate that PPARgamma binding to genomic target sites and recruitment of the mediator component TRAP220/MED1/PBP/DRIP205 is not affected by the deletion of the A/B-domain. By contrast, the PPARgamma-mediated CBP and p300 recruitment to A/B-domain dependent target genes is abolished by deletion of the A/B-domain. These results indicate that the A/B-domain of PPARgamma2 is specifically involved in the recruitment or stabilization of CBP and p300 containing co-factor complexes to a subset of target genes. Experiment Overall Design: Total RNA samples were obtained from three individual experiments in which NIH-CAR (i.e. a clonal cell line of NIH-3T3 fibroblasts stably expressing the coxsackie-adenovirus receptor (CARÎ1)) cells were transduced with adenovirus containing empty vector (AdEmpty) followed by addition of vehicle (DMSO) or transduced with adenovirus encoding either full length HA-tagged PPARg2 or A/B-domain deleted HA-PPARgCDE and treated with rosiglitazone.

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Intestinal Paneth cell differentiation relies on asymmetric regulation of Wnt signaling by Daam1/2

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