Cargando…
Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway
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-dime...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2009
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2779086/ https://www.ncbi.nlm.nih.gov/pubmed/19888210 http://dx.doi.org/10.1038/msb.2009.72 |
_version_ | 1782174335396478976 |
---|---|
author | Miller, Bryan W Lau, Garnet Grouios, Chris Mollica, Emanuela Barrios-Rodiles, Miriam Liu, Yongmei Datti, Alessandro Morris, Quaid Wrana, Jeffrey L Attisano, Liliana |
author_facet | Miller, Bryan W Lau, Garnet Grouios, Chris Mollica, Emanuela Barrios-Rodiles, Miriam Liu, Yongmei Datti, Alessandro Morris, Quaid Wrana, Jeffrey L Attisano, Liliana |
author_sort | Miller, Bryan W |
collection | PubMed |
description | 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 | Text |
id | pubmed-2779086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-27790862009-11-20 Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway Miller, Bryan W Lau, Garnet Grouios, Chris Mollica, Emanuela Barrios-Rodiles, Miriam Liu, Yongmei Datti, Alessandro Morris, Quaid Wrana, Jeffrey L Attisano, Liliana Mol Syst Biol Article 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. Nature Publishing Group 2009-10-13 /pmc/articles/PMC2779086/ /pubmed/19888210 http://dx.doi.org/10.1038/msb.2009.72 Text en Copyright © 2009, EMBO and Nature Publishing Group http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits distribution and reproduction in any medium, provided the original author and source are credited. This licence does not permit commercial exploitation or the creation of derivative works without specific permission. |
spellingShingle | Article Miller, Bryan W Lau, Garnet Grouios, Chris Mollica, Emanuela Barrios-Rodiles, Miriam Liu, Yongmei Datti, Alessandro Morris, Quaid Wrana, Jeffrey L Attisano, Liliana Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway |
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 | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2779086/ https://www.ncbi.nlm.nih.gov/pubmed/19888210 http://dx.doi.org/10.1038/msb.2009.72 |
work_keys_str_mv | AT millerbryanw applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT laugarnet applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT grouioschris applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT mollicaemanuela applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT barriosrodilesmiriam applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT liuyongmei applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT dattialessandro applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT morrisquaid applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT wranajeffreyl applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway AT attisanoliliana applicationofanintegratedphysicalandfunctionalscreeningapproachtoidentifyinhibitorsofthewntpathway |