Pax8 regulates proximal-tubule-specific identity via distal enhancer elements
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE300117 | GEO | 2026/07/16
REPOSITORIES: GEO
Similar Datasets
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In this submission, the PT-22 line was transduced with a Tet-on expression system linked to a hemagglutinin (HA)-tagged Pax8a transgene and a mixed population of successfully transduced cells selected. PAX8 was depleted from these modified PT-22 cells with tamoxifen, then rescued by inducing re-expression of Pax8a transgene with doxycycline. Gene expression was measured to determine gene targets altered by PAX8 re-expression.
2026-07-15 | GSE317090 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and gene expression was measured over time to identify likely activating and repressive targets of PAX8.
2026-07-15 | GSE317089 | GEO
Project description:Pax2 and Pax8 are homologous transcription factors required for kidney development and medullary urine concentration. However, their function in proximal tubule homeostasis and response to acute kidney injury is unknown. Mice with proximal tubules consisting of a mosaic of wild-type and Pax2/8 mutant proximal tubules cells were generated. Gene expression of mutant and wild-type proximal tubule cells was compared under homeostatic conditions using single-nucleus RNA sequencing.
2024-11-15 | GSE278289 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and Pax8 chromatin occupancy and histone marks (H3K4me3 and H3K27me3) were measured after 6 d using chromatin immunoprecipitation followed by sequencing (ChIP-seq).
2026-07-15 | GSE317088 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and chromatin accessibility was measured after 0, 3, and 24 d using Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq).
2026-07-15 | GSE317087 | GEO
Project description:We profiled SUMO1 and SUMO2 modified proteins in distal convoluted tubule (mpkDCT) as a precursor to targeted approaches to assess SUMO function. SUMO levels in cells following heat shock and treatment with deSUMOylation inhibitors were assessed.
2021-11-25 | PXD028507 | Pride
Project description:PBRM1 is the 2nd-most frequently inactivated gene in clear cell renal cell cancer (RCC) but the oncogenic mechanisms, and hence methods for correction, are unclear. PBRM1 is a subunit of the PBAF coactivator complex that transcription factors use to reposition nucleosomes (‘open chromatin’) for gene activation. We therefore looked for transcription factors that recruit endogenous PBRM1 in kidney lineage/RCC cells, as a waypoint to identifying pathways impacted by PBRM1 loss. Unbiased immunoprecipitation/mass-spectrometry analyses of the endogenous PBRM1 interactome in kidney lineage cells revealed PAX8, a master transcription factor essential for proximal tubule epithelial fates, as the major transcription factor recruiting PBRM1/PBAF. The reverse analyses of the PAX8 interactome confirmed recruitment specifically of PBRM1/PBAF, and not the functionally similar BAF coactivator complex. More conspicuous in the PAX8 hub in RCC cells, however, were several corepressors, e.g. DNMT1, which oppose coactivators to repress instead of activate genes. Accordingly, key PAX8 target genes, e.g., GATA3, LHX1, WT1, and ~1000 other downstream kidney epithelial genes, but not PAX8 or PAX2, demonstrated loss of the histone lysine 27 acetylation (H3K27ac) activation mark, increase in CpG methylation repression marks, and lower expression in RCC vs normal kidney cortex, with the greatest repression in cases with bi-allelic PBRM1 inactivation. PBRM1 re-introduction into RCC cells, or depletion of the corepressor DNMT1 using siRNA or a clinical drug decitabine, rebalanced composition and function of the PAX8 transcription factor hub to coactivators, to thereby activate terminal epithelial-fates in vitro and in vivo. In sum, PBRM1 loss in RCC cells skews coregulator composition of the PAX8 master transcription factor hub toward corepressors and repression instead of activation of the downstream terminal epithelial-program; this oncogenic action could be reversed by pharmacologic corepressor depletion/inhibition.
2022-02-16 | PXD020544 | Pride
Project description:Global gene expression in the primary cultured mouse kidney proximal tubule cells treated either DMSO or 1uM GW4064 (a FXR agonist) was compared. Results provide insight into mechanisms underlying effects of FXR activation on gene expression in mouse kidney proximal tubule cells. Male C57/BJ mice aged 6 weeks were sacrificed under anesthesia and kidney proximal tubule cells were cultured until confluent. Cells were treated with either GW4064 (1uM) or equal amount of DMSO and incubated for 24 hours. 4 total RNA samples per group were analyzed and gene expression was compared between the groups.
2015-06-26 | E-GEOD-70296 | biostudies-arrayexpress
Project description:Distal convoluted tubule (DCT) cells display a remarkable ability to adapt to the dietary potassium intake. We applied single-nucleus RNA-seq to map the plasticity of enriched mouse DCT cells in response to dietary potassium restriction
2023-12-31 | GSE228367 | GEO
Project description:Background: The mammalian kidneys maintain salt and water homeostasis for proper electrolyte balance and hydration. As the glomerular filtrate passes through the nephron and into the renal medulla, electrolytes, water, and urea are reabsorbed through the concerted actions of solute carrier channels and aquaporins located at various positions along the nephron and in the outer and inner medulla. Renal epithelial cells develop from Pax2 positive proliferating stem cells that suppress Pax2 expression once differentiated into mature proximal and distal tubules, but continue to express the related Pax8 protein. The collecting tubules and renal medulla are derived from a Pax2 positive ureteric bud epithelia that continue to express Pax2 and Pax8 in adult kidneys. Despite the necessity for Pax2 in renal development, functions for Pax2 or Pax8 in adult renal epithelia have not been established. Methods: In this report, we deleted either Pax2, Pax8, or both genes in adult mice and examined the phenotypes and changes in gene expression patterns. The mechanism of Pax8 mediated activation of potential target genes was described in inner medullary collecting duct cells. Results: Mice with induced deletions of both Pax2 and Pax8 exhibit severe polyuria that can be attributed to significant changes in the expression of solute carriers, such as the urea transporter UTA1, and aquaporins within the inner and outer medulla. Furthermore, Pax8 expression is induced by high salt in collecting duct cells and activates the UTA1 gene by recruiting a histone methyltransferase complex to the promoter. Conclusions: These data uncover novel functions for Pax proteins, in adult renal epithelia, that are essential for retaining water and concentrating urine.
2019-12-05 | GSE141455 | GEO