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A transcriptional response to Wnt protein in human embryonic carcinoma cells.

Willert J, Epping M, Pollack JR, Brown PO, Nusse R - BMC Dev. Biol. (2002)

Bottom Line: The promoters of nearly all the target genes we identified have putative TCF binding sites, and we show that the TCF binding site is required for induction of Follistatin.Several of the target genes have a cooperative response to a combination of Wnt and BMP.Wnt signaling activates genes that promote stem cell fate and inhibit cellular differentiation and regulates a remarkable number of genes involved in its own signaling system.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Developmental Biology, Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305 USA. jennifer.willert@cox.net

ABSTRACT

Background: Wnt signaling is implicated in many developmental decisions, including stem cell control, as well as in cancer. There are relatively few target genes known of the Wnt pathway.

Results: We have identified target genes of Wnt signaling using microarray technology and human embryonic carcinoma cells stimulated with active Wnt protein. The ~50 genes upregulated early after Wnt addition include the previously known Wnt targets Cyclin D1, MYC, ID2 and betaTRCP. The newly identified targets, which include MSX1, MSX2, Nucleophosmin, Follistatin, TLE/Groucho, Ubc4/5E2, CBP/P300, Frizzled and REST/NRSF, have important implications for understanding the roles of Wnts in development and cancer. The protein synthesis inhibitor cycloheximide blocks induction by Wnt, consistent with a requirement for newly synthesized beta-catenin protein prior to target gene activation. The promoters of nearly all the target genes we identified have putative TCF binding sites, and we show that the TCF binding site is required for induction of Follistatin. Several of the target genes have a cooperative response to a combination of Wnt and BMP.

Conclusions: Wnt signaling activates genes that promote stem cell fate and inhibit cellular differentiation and regulates a remarkable number of genes involved in its own signaling system.

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Related in: MedlinePlus

Microarray cluster analysis demonstrating increased expression of Wnt-3A responsive genes, including MSX1, MSX2, ID2, Versican, NPM, Frizzled-7, TLE/Groucho, Cyclin D1, and MYC. NCCIT cells were exposed to control conditioned medium (CCM) and Wnt-3A CM [8] for 4 hours. Shown here are 5 separate experiments. Note also that MSX 2 is included on this particular array in duplicate which provides an internal check on the precision of the measurements. Additional array experiments led to the identification of Follistatin and REST/NRSF as elevated genes (which were confirmed in Figure 2A). Brightest red corresponds to ratio of = 2:1 and dimmer red corresponds to ratio of 2:1. The full data set will be available at the Wnt homepage  and the Stanford Microarray database
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Figure 1: Microarray cluster analysis demonstrating increased expression of Wnt-3A responsive genes, including MSX1, MSX2, ID2, Versican, NPM, Frizzled-7, TLE/Groucho, Cyclin D1, and MYC. NCCIT cells were exposed to control conditioned medium (CCM) and Wnt-3A CM [8] for 4 hours. Shown here are 5 separate experiments. Note also that MSX 2 is included on this particular array in duplicate which provides an internal check on the precision of the measurements. Additional array experiments led to the identification of Follistatin and REST/NRSF as elevated genes (which were confirmed in Figure 2A). Brightest red corresponds to ratio of = 2:1 and dimmer red corresponds to ratio of 2:1. The full data set will be available at the Wnt homepage and the Stanford Microarray database

Mentions: To identify target genes, we applied Wnt-3A CM or CCM to the NCCIT cells and isolated RNA at several time points. We performed differential hybridization to microarray slides containing approximately 23,000 spots of human cDNAs, using RNA from CCM-exposed cells as a reference. Figure 1 shows a cluster analysis of the hybridization results. We found approximately 50 genes that were upregulated between 2 and 10 fold by Wnt-3A CM whereas a few genes were repressed, i.e. expressed at lower levels in the Wnt-3A-treated cells. The latter group consisted mostly of ESTs of unknown genes and has yet to be characterized.


A transcriptional response to Wnt protein in human embryonic carcinoma cells.

Willert J, Epping M, Pollack JR, Brown PO, Nusse R - BMC Dev. Biol. (2002)

Microarray cluster analysis demonstrating increased expression of Wnt-3A responsive genes, including MSX1, MSX2, ID2, Versican, NPM, Frizzled-7, TLE/Groucho, Cyclin D1, and MYC. NCCIT cells were exposed to control conditioned medium (CCM) and Wnt-3A CM [8] for 4 hours. Shown here are 5 separate experiments. Note also that MSX 2 is included on this particular array in duplicate which provides an internal check on the precision of the measurements. Additional array experiments led to the identification of Follistatin and REST/NRSF as elevated genes (which were confirmed in Figure 2A). Brightest red corresponds to ratio of = 2:1 and dimmer red corresponds to ratio of 2:1. The full data set will be available at the Wnt homepage  and the Stanford Microarray database
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC117803&req=5

Figure 1: Microarray cluster analysis demonstrating increased expression of Wnt-3A responsive genes, including MSX1, MSX2, ID2, Versican, NPM, Frizzled-7, TLE/Groucho, Cyclin D1, and MYC. NCCIT cells were exposed to control conditioned medium (CCM) and Wnt-3A CM [8] for 4 hours. Shown here are 5 separate experiments. Note also that MSX 2 is included on this particular array in duplicate which provides an internal check on the precision of the measurements. Additional array experiments led to the identification of Follistatin and REST/NRSF as elevated genes (which were confirmed in Figure 2A). Brightest red corresponds to ratio of = 2:1 and dimmer red corresponds to ratio of 2:1. The full data set will be available at the Wnt homepage and the Stanford Microarray database
Mentions: To identify target genes, we applied Wnt-3A CM or CCM to the NCCIT cells and isolated RNA at several time points. We performed differential hybridization to microarray slides containing approximately 23,000 spots of human cDNAs, using RNA from CCM-exposed cells as a reference. Figure 1 shows a cluster analysis of the hybridization results. We found approximately 50 genes that were upregulated between 2 and 10 fold by Wnt-3A CM whereas a few genes were repressed, i.e. expressed at lower levels in the Wnt-3A-treated cells. The latter group consisted mostly of ESTs of unknown genes and has yet to be characterized.

Bottom Line: The promoters of nearly all the target genes we identified have putative TCF binding sites, and we show that the TCF binding site is required for induction of Follistatin.Several of the target genes have a cooperative response to a combination of Wnt and BMP.Wnt signaling activates genes that promote stem cell fate and inhibit cellular differentiation and regulates a remarkable number of genes involved in its own signaling system.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Developmental Biology, Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305 USA. jennifer.willert@cox.net

ABSTRACT

Background: Wnt signaling is implicated in many developmental decisions, including stem cell control, as well as in cancer. There are relatively few target genes known of the Wnt pathway.

Results: We have identified target genes of Wnt signaling using microarray technology and human embryonic carcinoma cells stimulated with active Wnt protein. The ~50 genes upregulated early after Wnt addition include the previously known Wnt targets Cyclin D1, MYC, ID2 and betaTRCP. The newly identified targets, which include MSX1, MSX2, Nucleophosmin, Follistatin, TLE/Groucho, Ubc4/5E2, CBP/P300, Frizzled and REST/NRSF, have important implications for understanding the roles of Wnts in development and cancer. The protein synthesis inhibitor cycloheximide blocks induction by Wnt, consistent with a requirement for newly synthesized beta-catenin protein prior to target gene activation. The promoters of nearly all the target genes we identified have putative TCF binding sites, and we show that the TCF binding site is required for induction of Follistatin. Several of the target genes have a cooperative response to a combination of Wnt and BMP.

Conclusions: Wnt signaling activates genes that promote stem cell fate and inhibit cellular differentiation and regulates a remarkable number of genes involved in its own signaling system.

Show MeSH
Related in: MedlinePlus