Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2

<p>Abstract</p> <p>Background</p> <p>Wnt signaling is mediated through 1) the beta-catenin dependent canonical pathway and, 2) the beta-catenin independent pathways. Multiple receptors, including Fzds, Lrps, Ror2 and Ryk, are involved in Wnt signaling. Ror2 is a single-span transmembrane receptor-tyrosine kinase (RTK). The functions of Ror2 in mediating the non-canonical Wnt signaling have been well established. The role of Ror2 in canonical Wnt signaling is not fully understood.</p> <p>Results</p> <p>Here we report that Ror2 also positively modulates Wnt3a-activated canonical signaling in a lung carcinoma, H441 cell line. This activity of Ror2 is dependent on cooperative interactions with Fzd2 but not Fzd7. In addition, Ror2-mediated enhancement of canonical signaling requires the extracellular CRD, but not the intracellular PRD domain of Ror2. We further provide evidence that the positive effect of Ror2 on canonical Wnt signaling is inhibited by Dkk1 and Krm1 suggesting that Ror2 enhances an Lrp-dependent STF response.</p> <p>Conclusion</p> <p>The current study demonstrates the function of Ror2 in modulating canonical Wnt signaling. These findings support a functional scheme whereby regulation of Wnt signaling is achieved by cooperative functions of multiple mediators.</p

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-6

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>ne-rich domain; TM: transmembrane domain; Tyr-Kinase: tyrosine-kinase domain; S/T: serine and thronine rich domain; PRD: proline-rich domain

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-7

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>-type and mutant derivatives, and then treated with L-CM or Wnt3aCM. Equal amounts of DNA and conditioned media were used in each assay. Luciferase values were normalized to beta-galactosidase values for transfection efficiency. Data shown are the average ratios of normalized luciferase values in Wnt3aCM-treated cells over that of L-CM-treated cells. * indicates p < 0.05 when compared with data of bar 2. *** indicates p < 0.05 when compared with data of bar 3. indicates p < 0.05 when compared with data of bar 1. indicates p < 0.05 when compared with data of bar 8

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-2

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>, or Fzd7. An equal quantity of control vector was used as negative control. Twenty-four hours after transfection, cells were treated with L-CM and Wnt3aCM. Two dilutions of Wnt3aCM (1/8 or 1/3) were applied. Equal amounts of conditioned media were used in each assay. Luciferase values representing STF activities were normalized to beta-galactosidase for transfection efficiency. Values of the assays treated with L-CM alone were adjusted to unity to normalize all other experimental values in the same group. * indicates p < 0.05 when compared with data of bar 2. indicates p < 0.05 when compared with data of bar 3

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-1

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>human lung tissue was also determined. Expected size of each PCR product was listed in the table. Both Lrp5 and Lrp6 are expressed in A549 and H441 cells. Of the two Ror family members, only Ror1 is detectable

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-4

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>ts with or without Ror2WT plasmid and then treated with conditioned media, L-CM or Wnt3aCM. Equal amounts of DNA and conditioned media were used in each assay. Luciferase values were normalized to beta-galactosidase values for transfection efficiency. Values of the assays treated with L-CM alone (empty bars) were adjusted to unity to normalize all other experimental values in the same group. * indicates p < 0.05

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-11

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>ct, and then treated with L-CM or Wnt3aCM. Equal amounts of DNA and conditioned media were used in each assay. Luciferase values were normalized to beta-galactosidase for transfection efficiency. Values of the assay transfected with control vector and treated with L-CM alone (empty bars) were adjusted to unity to normalize all other experimental values in the same group. * indicates p < 0.05

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-5

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>es of Ror2 wild-type (Ror2WT) plasmid, and then treated with conditioned media, L-CM or Wnt3aCM. Equal amounts of DNA and conditioned media were used in each assay. Values of the assays were normalized as in Fig. 5. * indicates p < 0.05 when compared with data of bar 4. indicates p < 0.05 when compared with data of bar 2

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-12

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p> the control vector (-Ror2). Twenty four hours after transfection, cells were treated with L-CM, Wnt3aCM or Wnt5aCM. Wnt3aCM and Wnt5aCM were used at 1/8 and 1/4 dilutions, respectively. Equal amount of conditioned media was used in each assay. Luciferase values representing the STF activities were normalized to beta-galactosidase for transfection efficiency. Values of the assays treated with L-CM alone (#1 and #5) were adjusted to unity to normalize all other experimental values in the same group. CM: conditioned medium. * indicates p < 0.05

Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2-0

<p><b>Copyright information:</b></p><p>Taken from "Ror2 modulates the canonical Wnt signaling in lung epithelial cells through cooperation with Fzd2"</p><p>http://www.biomedcentral.com/1471-2199/9/11</p><p>BMC Molecular Biology 2008;9():11-11.</p><p>Published online 23 Jan 2008</p><p>PMCID:PMC2254434.</p><p></p>r the control vector (-Ror2). Twenty four hours after transfection, cells were treated with L-CM, Wnt3aCM or Wnt5aCM. Wnt3aCM and Wnt5aCM were used at 1/8 and 1/4 dilutions, respectively. Equal amount of conditioned media was used in each assay. Luciferase values representing the STF activities were normalized to beta-galactosidase for transfection efficiency. Values of the assays treated with L-CM alone (#1 and #5) were adjusted to unity to normalize all other experimental values in the same group. CM: conditioned medium. * indicates p < 0.05