Heterarchy of transcription factors driving basal and luminal cell phenotypes in human urothelium

Fishwick, Carl, Higgins, Janet, Percival-Alwyn, Lawrence, Hustler, Arianna, Pearson, Joanna, Bastkowski, Sarah, Moxon, Simon, Swarbreck, David, Greenman, Chris D. and Southgate, Jennifer (2017) Heterarchy of transcription factors driving basal and luminal cell phenotypes in human urothelium. Cell Death and Differentiation, 24. pp. 809-818. ISSN 1350-9047

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Abstract

Cell differentiation is effected by complex networks of transcription factors that co-ordinate re-organisation of the chromatin landscape. The hierarchies of these relationships can be difficult to dissect. During in vitro differentiation of normal human uro-epithelial cells, formaldehyde-assisted isolation of regulatory elements (FAIRE-seq) and RNA-seq were used to identify alterations in chromatin accessibility and gene expression changes following activation of the nuclear receptor PPARG as a differentiation-initiating event. Regions of chromatin identified by FAIRE-seq as having altered accessibility during differentiation were found to be enriched with sequence-specific binding motifs for transcription factors predicted to be involved in driving basal and differentiated urothelial cell phenotypes, including FOXA1, P63, GRHL2, CTCF and GATA3. In addition, co-occurrence of GATA3 motifs was observed within sub-sets of differentiation-specific peaks containing P63 or FOXA1 after induction of differentiation. Changes in abundance of GRHL2, GATA3, and P63 were observed in immunoblots of chromatin-enriched extracts. Transient siRNA knockdown of P63 revealed that P63 favoured a basal-like phenotype by inhibiting differentiation and promoting expression of basal marker genes. GATA3 siRNA prevented differentiation-associated downregulation of P63 protein and transcript, and demonstrated positive feedback of GATA3 on PPARG transcript, but showed no effect on FOXA1 transcript or protein expression. This approach indicates that as a transcriptionally-regulated programme, urothelial differentiation operates as a heterarchy wherein GATA3 is able to co-operate with FOXA1 to drive expression of luminal marker genes, but that P63 has potential to transrepress expression of the same genes.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
Faculty of Science > School of Computing Sciences
Depositing User: Pure Connector
Date Deposited: 13 Jan 2017 00:06
Last Modified: 22 Jul 2020 23:34
URI: https://ueaeprints.uea.ac.uk/id/eprint/62032
DOI: 10.1038/cdd.2017.10

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