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Profiling ERRb-target genes in mouse ES cells

ABSTRACT: Profiling ERRb-target genes in mouse ES cells

PROVIDER: PRJNA691464 | ENA |

REPOSITORIES: ENA

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Profiling ERRb-target genes in mouse ES cells

Project description:We found ERRb activates the transcription through two molecular pathways, which are interaction to AF2-region binding PGC-1a/cofactors and general transcription factor TFIIH. To analyze the significance of these pathways in cells, we first identified ERRb-target genes in ES cells. Estrogen-related receptors (ERRa/b/g) are orphan nuclear receptors that function in a number of energy-demanding physiological processes, as well as in development and stem cell maintenance, but mechanisms underlying target gene activation are largely unknown. Here, reconstituted biochemical assays that manifest ERR-dependent transcription have revealed two complementary mechanisms. On chromatin templates ERR-dependent transcription is dependent on interactions with cell-specific coactivator PGC-1a, which in turn recruits the ubiquitous p300 and MED1/Mediator coactivators. The N-terminal half of PGC-1a is necessary and sufficient for these interactions and for transcription both in vitro and in vivo. On DNA templates, ERRs activate transcription with just the normal complement of general initiation factors in a manner dependent upon interaction of the ERR DNA-binding domain with the p52 subunit of initiation factor TFIIH. Importantly, the PGC-1a and TFIIH interactions are both essential for ERRb/g functions in maintaining embryonic stem cell pluripotency through regulation of target gene transcription.

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HOXB4 target genes in ES cell-derived embryoid bodies (EBs)

Project description:To unravel the molecular mechanism by which HOXB4 promotes the expansion of early hematopoietic progenitors within differentiating ES cells, we analzed the gene expression profiles of embryoid bodies (EBs) in which transcription of HOXB4 had been induced or not induced. A substantial number of the identified HOXB4 target genes are involved in signaling pathways important for controlling self-renewal, maintenance and differentiation of stem cells. Furthermore, we demonstrate that HOXB4 activity and FGF-signaling are intertwined. HOXB4-mediated expansion of ES cell-derived early progenitors was enhanced by specific and complete inhibition of FGF-receptors. In contrast, the expanding activity of HOXB4 on hematopoietic progenitors in day4-6 embryoid bodies was blunted in the presence of basic FGF (FGF2) indicating a dominant negative effect of FGF-signaling on the earliest hematopoietic cells. Taken together, we show that modulation of FGF signaling is an essential feature of HOXB4 activity in the context of embryonic hematopoiesis. Keywords: plus/minus induction of HOXB4 gene expression by treatment with doxycycline (Dox)

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Project description:Human ES cells respond to activation of the BMP and WNT signaling by upregulating target genes. A 4h time-point following signaling factor stimulation was chosen to reveal immediate-early induced genes which are likely to be direct targets.

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