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P300 proximity labeling in human iPSCs

ABSTRACT: This dataset presents the results of proximity-dependent biotinylation targeting the transcriptional coactivator p300 in human induced pluripotent stem cells (hiPSCs). We endogenously tagged p300 with the biotin ligase miniTurboID, to enable labeling of proximal proteins and thus proteins locatd at enhancers. hiPSCs were cultured in two different conditions, either StemFit or mTeSR medium, to assess how media composition influences the p300 proximal proteome. Following biotin labeling, proteins were enriched via streptavidin pulldown and analyzed by LC-MS/MS.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell, Hipsc Cell

SUBMITTER: S Stelloo

LAB HEAD: Prof. Michiel

PROVIDER: PXD067912 | Pride | 2026-08-07

REPOSITORIES: Pride

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			Action	DRS
	20250227_Astral_AUR_SA_iPSCs_WT_mTSER_1.raw	Raw
	20250227_Astral_AUR_SA_iPSCs_WT_mTSER_2.raw	Raw
	20250227_Astral_AUR_SA_iPSCs_WT_mTSER_3.raw	Raw
	20250227_Astral_AUR_SA_iPSCs_WT_stemFit_7.raw	Raw
	20250227_Astral_AUR_SA_iPSCs_WT_stemFit_8.raw	Raw

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Project description:The transcriptional regulation of drug-metabolizing enzymes and transporters (here collectively referred to as DMEs) in the developing proximal tubule is not well understood. As in the liver, DME regulation in the PT may be mediated through nuclear receptors which are thought to “sense” deviations from homeostasis by being activated by ligands, some of which are handled by DMEs, including drug transporters. Systems analysis of transcriptomic data during kidney development predicted a set of upstream transcription factors, including Hnf4a and Hnf1a, as well as Nr3c1 (Gr), Nfe2l2 (Nrf2), Ppara, and Tp53. Motif analysis of cis-regulatory further suggested that Hnf4a and Hnf1a are the main transcriptional regulators in the PT. Available expression data from tissue-specific Hnf4a KO tissues revealed that distinct subsets of DMEs were regulated by Hnf4a in a tissue-specific manner. ChIP-seq was performed to characterize the PT-specific binding sites of Hnf4a in rat kidneys at three developmental stages (prenatal, immature, adult), which further supported a major role for Hnf4a in regulating PT gene expression, including DMEs. In ex vivo kidney organ culture, an antagonist of Hnf4a (but not a similar inactive compound) led to predicted changes in DME expression, including among others Fmo1, Cyp2d2, Cyp2d4, Nqo2, as well as organic cation transporters and organic anion transporters Slc22a1(Oct1), Slc22a2 (Oct2), Slc22a6 (Oat1), Slc22a8(Oat3), and Slc47a1(Mate1). Conversely, overexpression of Hnf1a and Hnf4a in primary mouse embryonic fibroblasts (MEFs), sometimes considered a surrogate for mesenchymal stem cells, induced expression of several of these proximal tubule DMEs, as well as epithelial markers and a PT-specific brush border marker Ggt1. These cells had organic anion transporter function. Taken together, the data strongly supports a critical role for HNF4a and Hnf1a in the tissue-specific regulation of drug handling and differentiation toward a PT cellular identity. Hnf4a binding was examined in rat kidneys at three timepoints (E20, P13 and Adult) and p300 binding was examined in adult rat kidney cortex tissue using ChIP-seq. Four corresponding input DNA samples were used as controls for peak calling.

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P300 proximity labeling in human iPSCs

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