Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway
Abstract Large‐scale proteomic approaches have been used to study signaling pathways. However, identification of biologically relevant hits from a single screen remains challenging due to limitations inherent in each individual approach. To overcome these limitations, we implemented an integrated, m...
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| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2009-10-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.1038/msb.2009.72 |
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| _version_ | 1849225795723067392 |
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| author | Bryan W Miller Garnet Lau Chris Grouios Emanuela Mollica Miriam Barrios‐Rodiles Yongmei Liu Alessandro Datti Quaid Morris Jeffrey L Wrana Liliana Attisano |
| author_facet | Bryan W Miller Garnet Lau Chris Grouios Emanuela Mollica Miriam Barrios‐Rodiles Yongmei Liu Alessandro Datti Quaid Morris Jeffrey L Wrana Liliana Attisano |
| author_sort | Bryan W Miller |
| collection | DOAJ |
| description | Abstract Large‐scale proteomic approaches have been used to study signaling pathways. However, identification of biologically relevant hits from a single screen remains challenging due to limitations inherent in each individual approach. To overcome these limitations, we implemented an integrated, multi‐dimensional approach and used it to identify Wnt pathway modulators. The LUMIER protein–protein interaction mapping method was used in conjunction with two functional screens that examined the effect of overexpression and siRNA‐mediated gene knockdown on Wnt signaling. Meta‐analysis of the three data sets yielded a combined pathway score (CPS) for each tested component, a value reflecting the likelihood that an individual protein is a Wnt pathway regulator. We characterized the role of two proteins with high CPSs, Ube2m and Nkd1. We show that Ube2m interacts with and modulates β‐catenin stability, and that the antagonistic effect of Nkd1 on Wnt signaling requires interaction with Axin, itself a negative pathway regulator. Thus, integrated physical and functional mapping in mammalian cells can identify signaling components with high confidence and provides unanticipated insights into pathway regulators. |
| format | Article |
| id | doaj-art-c93cae2ed5fc49f2a5aad7c0004c5e49 |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2009-10-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-c93cae2ed5fc49f2a5aad7c0004c5e492025-08-24T11:59:09ZengSpringer NatureMolecular Systems Biology1744-42922009-10-015111310.1038/msb.2009.72Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathwayBryan W Miller0Garnet Lau1Chris Grouios2Emanuela Mollica3Miriam Barrios‐Rodiles4Yongmei Liu5Alessandro Datti6Quaid Morris7Jeffrey L Wrana8Liliana Attisano9Department of Biochemistry, University of TorontoDepartment of Biochemistry, University of TorontoBanting and Best Department of Medical Research, University of TorontoDepartment of Biochemistry, University of TorontoCentre for Systems Biology, Samuel Lunenfeld Research Institute, Mount Sinai HospitalCentre for Systems Biology, Samuel Lunenfeld Research Institute, Mount Sinai HospitalSamuel Lunenfeld Research Institute, Mount Sinai HospitalDonnelly Centre for Cellular and Biomolecular Research, University of TorontoCentre for Systems Biology, Samuel Lunenfeld Research Institute, Mount Sinai HospitalDepartment of Biochemistry, University of TorontoAbstract Large‐scale proteomic approaches have been used to study signaling pathways. However, identification of biologically relevant hits from a single screen remains challenging due to limitations inherent in each individual approach. To overcome these limitations, we implemented an integrated, multi‐dimensional approach and used it to identify Wnt pathway modulators. The LUMIER protein–protein interaction mapping method was used in conjunction with two functional screens that examined the effect of overexpression and siRNA‐mediated gene knockdown on Wnt signaling. Meta‐analysis of the three data sets yielded a combined pathway score (CPS) for each tested component, a value reflecting the likelihood that an individual protein is a Wnt pathway regulator. We characterized the role of two proteins with high CPSs, Ube2m and Nkd1. We show that Ube2m interacts with and modulates β‐catenin stability, and that the antagonistic effect of Nkd1 on Wnt signaling requires interaction with Axin, itself a negative pathway regulator. Thus, integrated physical and functional mapping in mammalian cells can identify signaling components with high confidence and provides unanticipated insights into pathway regulators.https://doi.org/10.1038/msb.2009.72cancer biologysignal transductionsystems biology |
| spellingShingle | Bryan W Miller Garnet Lau Chris Grouios Emanuela Mollica Miriam Barrios‐Rodiles Yongmei Liu Alessandro Datti Quaid Morris Jeffrey L Wrana Liliana Attisano Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway Molecular Systems Biology cancer biology signal transduction systems biology |
| title | Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
| title_full | Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
| title_fullStr | Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
| title_full_unstemmed | Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
| title_short | Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
| title_sort | application of an integrated physical and functional screening approach to identify inhibitors of the wnt pathway |
| topic | cancer biology signal transduction systems biology |
| url | https://doi.org/10.1038/msb.2009.72 |
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