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Abstract: PO1104

Bayesian Identification of Transcription Factors That Regulate Aqp2 Transcription

Session Information

Category: Fluid, Electrolyte, and Acid-Base Disorders

  • 901 Fluid, Electrolyte, and Acid-Base Disorders: Basic

Authors

  • Kikuchi, Hiroaki, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States
  • Jung, Hyun Jun, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States
  • Knepper, Mark A., National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States
Background

Renal collecting duct and connecting tubule cells selectively express the water channel aquaporin-2 (AQP2) and Aqp2 gene transcription is strongly regulated by vasopressin. However, the transcription factors (TFs) responsible for regulation of expression of AQP2 remain largely unknown. Here, we used Bayesian data integration techniques to identify these TFs.

Methods

The general strategy is to use Bayes’ Rule to integrate several -omic datasets to stratify a curated list of 1344 TFs present in the mouse genome with regard to probability of regulating Aqp2 gene transcription. To prioritize TFs, existing proteomic and transcriptomic data, ATAC-Seq, histone H3K27-acetylation ChIP-Seq, and RNA-polymerase II ChIP-Seq data are used. Beyond this, we use additional -omic datasets to prioritize TFs that are regulated by vasopressin. Finally, we carried out new RNA-Seq experiments mapping the time course of vasopressin induced changes in the transcriptome of mouse mpkCCD cells to further prioritize TFs that change in tandem with AQP2.

Results

The analysis identified 17 TFs out of 1344 in the mouse genome that are most likely to be involved in regulation of Aqp2 gene transcription. These TFs included eight that have been proposed in prior studies to play a role in Aqp2 regulation, viz. Cebpb, Elf1, Elf3, Ets1, Jun, Junb, Nfkb1, and Sp1. The remaining nine represent new candidates for future studies (Atf1, Irf3, Klf5, Klf6, Mef2d, Nfyb, Nr2f6, Stat3, Nr4a1). The RNA-Seq time course experiments in mpkCCD cells showed a rapid increase in Aqp2 mRNA, within 3 hour of vasopressin exposure. This response was matched by an equally rapid increase in the abundance of the mRNA coding for Cebpb, which we have shown by ChIP-seq studies to bind downstream from the Aqp2 gene.

Conclusion

The Bayesian analysis has identified the TFs most likely to bind to Aqp2 cis-regulatory elements and likely to be regulated by vasopressin stimulation, providing a roadmap for future studies to understand regulation of Aqp2 gene expression.

Funding

  • Other NIH Support