Yan, an ETS‐domain transcription factor, negatively modulates the Wingless pathway in the <i>Drosophila</i> eye

Olson, Emily R; Pancratov, Raluca; Chatterjee, Satabdi; Changkakoty, Binita; Pervaiz, Zeeshan; DasGupta, Ramanuj

doi:10.1038/embor.2011.159

Cited by 37 publications

(55 citation statements)

References 29 publications

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“…To further test the hypothesis that Tnks promotes Wingless signaling in the adult intestine, we analyzed reporters for the Wingless pathway target genes naked (nkd) and frizzled 3 ( fz3) (Olson et al, 2011;Sato et al, 1999;Sivasankaran et al, 2000;Zeng et al, 2000). Previous studies revealed that the nkd-lacZ and the fz3-RFP reporters are expressed in overlapping regional gradients along the length of the adult intestine and indicated that Wingless pathway activity is highest at all the major boundaries that exist between compartments (Fig.…”

Section: Gfp (Green) and Ebs By Su(h)-lacz (Magenta) (G-i) Tnks Mutamentioning

confidence: 99%

“…Other stocks used were: esg-Gal4, UAS-GFP (Micchelli and Perrimon, 2006), esg K606 (Micchelli and Perrimon, 2006), Su(H)GBE-lacZ (Micchelli and Perrimon, 2006), fz3-RFP (Olson et al, 2011), nkd-lacZ (Zeng et al, 2000), 10× stat-GFP (Bach et al, 2007), hs-flp, UAS-mCD8:GFP; tub-Gal4, FRT82B, tub-Gal80 (Li et al, 2013a), hs-flp, tub-gal4, UAS-dsRed; FRT82B, tub-gal80 (Guo et al, 2013), hs-flp, tubGal80, FRT19A; tub-Gal4, UAS-mCD8:GFP (Choi et al, 2011), FRT82B (BDSC), 82B pygo S123 (Thompson et al, 2002), dsh V26 FRT19A (Klingensmith et al, 1994;Wehrli and Tomlinson, 1998), tub>Flag-Axin; Axin s044230 (Peterson-Nedry et al, 2008), Axin s044230 (Hamada et al, 1999), BAC Axin-V5 integrated at the VK30 site (PBac{y[+]-attP-9A}VK00030) (Gerlach et al, 2014), Myo1A-Gal4 (Karpowicz et al, 2013), Myo1A-Gal4, tub-Gal80 ts , UAS-GFP (Jiang et al, 2009), esg-Gal4, tub-Gal80 ts , UAS-GFP (Micchelli and Perrimon, 2006), UAS-Tnks RNAi#1 (VDRC#106238), UAS-Tnks RNAi#2 (Tnks-RNAi-2) (Feng et al, 2014), UAS-dsh RNAi (VDRC#31306), UAS-upd3 RNAi#1 (VDRC#106869), UAS-upd3 RNAi#2 (VDRC#27136), UAS-rpr3-5/TM3. Canton S flies were used as controls, and all crosses were performed at 25°C unless otherwise indicated.…”

Section: Flies and Geneticsmentioning

confidence: 99%

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The ADP-ribose polymerase Tankyrase regulates adult intestinal stem cell proliferation during homeostasis in Drosophila

et al. 2016

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Wnt/β-catenin signaling controls intestinal stem cell (ISC) proliferation, and is aberrantly activated in colorectal cancer. Inhibitors of the ADP-ribose polymerase Tankyrase (Tnks) have become lead therapeutic candidates for Wnt-driven cancers, following the recent discovery that Tnks targets Axin, a negative regulator of Wnt signaling, for proteolysis. Initial reports indicated that Tnks is important for Wnt pathway activation in cultured human cell lines. However, the requirement for Tnks in physiological settings has been less clear, as subsequent studies in mice, fish and flies suggested that Tnks was either entirely dispensable for Wntdependent processes in vivo, or alternatively, had tissue-specific roles. Here, using null alleles, we demonstrate that the regulation of Axin by the highly conserved Drosophila Tnks homolog is essential for the control of ISC proliferation. Furthermore, in the adult intestine, where activity of the Wingless pathway is graded and peaks at each compartmental boundary, Tnks is dispensable for signaling in regions where pathway activity is high, but essential where pathway activity is relatively low. Finally, as observed previously for Wingless pathway components, Tnks activity in absorptive enterocytes controls the proliferation of neighboring ISCs non-autonomously by regulating JAK/STAT signaling. These findings reveal the requirement for Tnks in the control of ISC proliferation and suggest an essential role in the amplification of Wnt signaling, with relevance for development, homeostasis and cancer.

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Section: Gfp (Green) and Ebs By Su(h)-lacz (Magenta) (G-i) Tnks Mutamentioning

confidence: 99%

Section: Flies and Geneticsmentioning

confidence: 99%

The ADP-ribose polymerase Tankyrase regulates adult intestinal stem cell proliferation during homeostasis in Drosophila

et al. 2016

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“…Development 140 (14) to specify the sensory wing margin bristles by activating Sens (Olson et al, 2011). Additionally, we used MARCM to assess whether misexpression of miR-310/13 could suppress the ectopic activation of Sens in clones of cells lacking WT Axin (supplementary material Fig.…”

Section: Research Articlementioning

confidence: 99%

The miR-310/13 cluster antagonizes β-catenin function in the regulation of germ and somatic cell differentiation in theDrosophilatestis

et al. 2013

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SUMMARYMicroRNAs (miRNAs) are regulators of global gene expression and function in a broad range of biological processes. Recent studies have suggested that miRNAs can function as tumor suppressors or oncogenes by modulating the activities of evolutionarily conserved signaling pathways that are commonly dysregulated in cancer. We report the identification of the miR-310 to miR-313 (miR-310/13) cluster as a novel antagonist of Wingless (Drosophila Wnt) pathway activity in a functional screen for Drosophila miRNAs. We demonstrate that miR-310/13 can modulate Armadillo (Arm; Drosophila β-catenin) expression and activity by directly targeting the 3ʹ-UTRs of arm and pangolin (Drosophila TCF) in vivo. Notably, the miR-310/13-deficient flies exhibit abnormal germ and somatic cell differentiation in the male gonad, which can be rescued by reducing Arm protein levels or activity. Our results implicate a previously unrecognized function for miR-310/13 in dampening the activity of Arm in early somatic and germline progenitor cells, whereby inappropriate/sustained activation of Arm-mediated signaling or cell adhesion may impact normal differentiation in the Drosophila male gonad. MATERIALS AND METHODS Cell culture and high-throughput screen (HTS)For the HTS, the Wg pathway was activated in Drosophila Clone 8 (Cl8) and S2R+ cells [grown as described by DasGupta et al. (DasGupta et al., 2005)] by introducing Axin double-stranded RNA (dsRNA), which resulted in a robust, ligand-independent activation of the Wg-responsive dTF12 reporter (DasGupta et al., 2005) (Fig. 1A). We screened a library of miRNA expression constructs [UAS-dsRED-pri-miR (Silver et al., 2007)] that consisted of 75 previously screened pri-miR constructs (Silver et al., 2007) plus 115 as yet unscreened pri-miR plasmids for their ability to suppress dTF12 activity downstream of the DC in this transcriptionally sensitized background. A total of 190 screen-ready plasmids were plated using a Janus MDT automated workstation (Perkin Elmer) in 5 µl aliquots as quadruplicates arranged in a quadrant on a set of three 384-well plates. Several quadrants of four replica wells were left empty for the addition of assay-specific controls. Axin dsRNA was generated using the Megascript kit (Applied Biosystems) using the following primers (5Ј-3Ј): forward TAATACGACTCACTATAGGGagaccaaacgccgcaccgctcgcc and reverse TAATACGACTCACTATAGGGagacaaaagccggtcgcccgtac (capital letters denote priming regions for T7 RNA polymerase).Cells were suspended at 20,000 cells/well for S2/S2R+ and 40,000 cells/well for Cl8. The dTF12-luciferase (TOP12-Ffl) reporter and Pol IIIRenilla luciferase (PolIII-RL) were utilized as described (DasGupta et al., 2005), with the addition of 0.01 µg actin-GAL4 and 0.1 µg Axin dsRNA, and transfected using the Effectene kit (Qiagen). Cells were incubated posttransfection for 5 days and luciferase levels assessed using the Promega Dual-Glo kit (Promega).For screen data analysis, Firefly luciferase activity values were normalized to those of Renilla luciferase for each...

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“…Other stocks used were Df(1)EDF7161 P(w+) [11A1;11B14] / Dp(1;Y)BSC1, y+ [10C1-2;11D3-8], aoslacZ W11 , E(spl)m8-lacZ, ds-lacZ 05124 , Dll-lacZ 01092 , dpplacZ BS3.0 , ptc-GAL4 559.1 , ptc-lacZ AT90 , vgBE-lacZ (Bloomington Drosophila Stock Center), Y hs-hid (a gift from Ruth Lehmann), fz3-RFP (Olson et al 2011), UAS-dTCF DN (van de Wetering et al 1997), UAS-CK1a::Flag, UAS-CG2577 (Zhang et al 2006), UAS-dco KD (Kinase Domain), dco 3 (Cho et al 2006). Stocks used to generate clones were: (1) y,w, eyFLP1, P[w+, arm-lacZ], FRT19A, (2) w, hsFLP 122 , P[w+, ubi-GFP], FRT19A, (3) y,w, hsFLP 122 , P[w+, ubi-RFP], FRT19A, (4) y,w, P[w+, ubi-GFP], M(1) RpS5a 2 , FRT19A; hsFLP 38 , (5) w, eyFLP1, tub-GAL80, FRT19A;; tub-GAL4, UAS-CD8::GFP, (6) w, hsFLP 122 , tub-GAL80, FRT19A;; tub-GAL4, UAS-CD8::GFP.…”

Section: Fly Stocks and Geneticsmentioning

confidence: 99%

“…However, the presence of eight CK1 family members in Drosophila and the difficulty of achieving both efficient and specific loss of function by RNAi have left open the possibility that other CK1 family members contribute to Arm degradation in vivo (Jia et al 2005;Zhang et al 2006). We tested this using an enhancer from the frizzled 3 (fz3) gene that is directly regulated by Wg through TCF binding sites (Olson et al 2011). fz3-RFP, which is normally expressed in response to high Wg signaling in the notum and to a lesser extent at the D/V boundary in the wing disc ( Figure 7A), was strongly induced in CK1a 8B12 clones (Figure 7B), indicating robust pathway activation.…”

Section: Ck1a Is Specifically Required For Wg Signalingmentioning

confidence: 99%

A Screen for X-Linked Mutations Affecting Drosophila Photoreceptor Differentiation Identifies Casein Kinase 1α as an Essential Negative Regulator of Wingless Signaling

et al. 2012

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The Wnt and Hedgehog signaling pathways are essential for normal development and are misregulated in cancer. The casein kinase family of serine/threonine kinases regulates both pathways at multiple levels. However, it has been difficult to determine whether individual members of this family have distinct functions in vivo, due to their overlapping substrate specificities. In Drosophila melanogaster, photoreceptor differentiation is induced by Hedgehog and inhibited by Wingless, providing a sensitive system in which to identify regulators of each pathway. We used a mosaic genetic screen in the Drosophila eye to identify mutations in genes on the X chromosome required for signal transduction. We recovered mutations affecting the transcriptional regulator CREB binding protein, the small GTPase dynamin, the cytoskeletal regulator Actin-related protein 2, and the protein kinase Casein kinase 1a. Consistent with its reported function in the b-Catenin degradation complex, Casein Kinase 1a mutant cells accumulate b-Catenin and ectopically induce Wingless target genes. In contrast to previous studies based on RNA interference, we could not detect any effect of the same Casein Kinase 1a mutation on Hedgehog signaling. We thus propose that Casein kinase 1a is essential to allow b-Catenin degradation and prevent inappropriate Wingless signaling, but its effects on the Hedgehog pathway are redundant with other Casein kinase 1 family members.

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Yan, an ETS‐domain transcription factor, negatively modulates the Wingless pathway in the Drosophila eye

Cited by 37 publications

References 29 publications

The ADP-ribose polymerase Tankyrase regulates adult intestinal stem cell proliferation during homeostasis in Drosophila

The ADP-ribose polymerase Tankyrase regulates adult intestinal stem cell proliferation during homeostasis in Drosophila

The miR-310/13 cluster antagonizes β-catenin function in the regulation of germ and somatic cell differentiation in theDrosophilatestis

A Screen for X-Linked Mutations Affecting Drosophila Photoreceptor Differentiation Identifies Casein Kinase 1α as an Essential Negative Regulator of Wingless Signaling

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