Wnt/β-catenin and LIF/Stat3 signaling pathways converge on Sp5 to promote mouse embryonic stem cell self-renewal

Ye, Shoudong; Zhang, Dongming; Cheng, Fei; Wilson, Daniel N.; Mackay, Jeffrey; He, Kan; Ban, Qian; Lv, Feng; Huang, Saifei; Liu, Dahai; Ying, Qi‐Long

doi:10.1242/jcs.177675

Cited by 48 publications

(43 citation statements)

References 41 publications

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“…SP5, which shares limited domain homology with the ubiquitous SP1 transcription factor (Supplementary Fig. 1h ), has previously been described as a WNT/β-catenin target gene 12 , 14 – 18 . Immunoblotting with an antibody directed against the amino-terminus of SP5 (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

The WNT target SP5 negatively regulates WNT transcriptional programs in human pluripotent stem cells

et al. 2017

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The WNT/β-catenin signaling pathway is a prominent player in many developmental processes, including gastrulation, anterior–posterior axis specification, organ and tissue development, and homeostasis. Here, we use human pluripotent stem cells (hPSCs) to study the dynamics of the transcriptional response to exogenous activation of the WNT pathway. We describe a mechanism involving the WNT target gene SP5 that leads to termination of the transcriptional program initiated by WNT signaling. Integration of gene expression profiles of wild-type and SP5 mutant cells with genome-wide SP5 binding events reveals that SP5 acts to diminish expression of genes previously activated by the WNT pathway. Furthermore, we show that activation of SP5 by WNT signaling is most robust in cells with developmental potential, such as stem cells. These findings indicate a mechanism by which the developmental WNT signaling pathway reins in expression of transcriptional programs.

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Section: Resultsmentioning

confidence: 99%

The WNT target SP5 negatively regulates WNT transcriptional programs in human pluripotent stem cells

et al. 2017

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“…Overlapping PCR was used to generate the Gsk3α-L195G, Gsk3α-K148R (kinase-dead) , Gsk3β-L132G, Gsk3β-K85R (kinase-dead) mutations. ES cells were transfected with PiggyBac vector (Ye et al, 2016) carrying various GSK3 mutants using Lipofectamine LTX (Invitrogen) according to the manufacturer’s instructions. Selection began one day after transfection by adding puromycin/hygromycin/blasticidin/zeocin and continued for one week or until the untransfected cells were all killed by the antibiotics in the untransfected control group.…”

Section: Star*methodsmentioning

confidence: 99%

A Chemical-Genetic Approach Reveals the Distinct Roles of GSK3α and GSK3β in Regulating Embryonic Stem Cell Fate

et al. 2017

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SUMMARY Glycogen synthase kinase 3 (GSK3) plays a central role in diverse cellular processes. GSK3 has two mammalian isozymes, GSK3α and GSK3β, whose functions remain ill-defined because of a lack of inhibitors that can distinguish between the two highly homologous isozymes. Here, we show that GSK3α and GSK3β can be selectively inhibited in mouse embryonic stem cells (ESCs) using a chemical-genetic approach. Selective inhibition of GSK3β is sufficient to maintain mouse ESC self-renewal, whereas GSK3α inhibition promotes mouse ESC differentiation toward neural lineages. Genome-wide transcriptional analysis reveals that GSK3α and GSK3β have distinct sets of downstream targets. Furthermore, selective inhibition of individual GSK3 isozymes yields distinct phenotypes from gene deletion, highlighting the power of the chemical-genetic approach in dissecting kinase catalytic functions from the protein’s scaffolding functions. Our study opens new avenues for defining GSK3 isozyme-specific functions in various cellular processes.

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“…Our previous report showed that Sp5 is a downstream target of STAT3 and overexpression of Sp5 is sufficient to maintain the undifferentiated state of mESCs in the absence of LIF [ 15 ]. Sp5, a member of the Sp1 family, is characterized by the presence of three typical zinc finger domains belonging to the specificity protein/Krϋppel-like factor (Sp/Klf) superfamily [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Sp5 induces the expression of Nanog to maintain mouse embryonic stem cell self-renewal

et al. 2017

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Activation of signal transducer and activator of transcription 3 (STAT3) by leukemia inhibitory factor (LIF) maintains mouse embryonic stem cell (mESC) self-renewal. Our previous study showed that trans-acting transcription factor 5 (Sp5), an LIF/STAT3 downstream target, supports mESC self-renewal. However, the mechanism by which Sp5 exerts these effects remains elusive. Here, we found that Nanog is a direct target of Sp5 and mediates the self-renewal-promoting effect of Sp5 in mESCs. Overexpression of Sp5 induced Nanog expression, while knockdown or knockout of Sp5 decreased the Nanog level. Moreover, chromatin immunoprecipitation (ChIP) assays showed that Sp5 directly bound to the Nanog promoter. Functional studies revealed that knockdown of Nanog eliminated the mESC self-renewal-promoting ability of Sp5. Finally, we demonstrated that the self-renewal-promoting function of Sp5 was largely dependent on its zinc finger domains. Taken together, our study provides unrecognized functions of Sp5 in mESCs and will expand our current understanding of the regulation of mESC pluripotency.

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Wnt/β-catenin and LIF/Stat3 signaling pathways converge on Sp5 to promote mouse embryonic stem cell self-renewal

Cited by 48 publications

References 41 publications

The WNT target SP5 negatively regulates WNT transcriptional programs in human pluripotent stem cells

The WNT target SP5 negatively regulates WNT transcriptional programs in human pluripotent stem cells

A Chemical-Genetic Approach Reveals the Distinct Roles of GSK3α and GSK3β in Regulating Embryonic Stem Cell Fate

Sp5 induces the expression of Nanog to maintain mouse embryonic stem cell self-renewal

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