Application of an integrated physical and functional screening approach to identify inhibitors of the Wnt pathway

Autor: Chris Grouios, Liliana Attisano, Emanuela Mollica, Quaid Morris, Jeffrey L. Wrana, Yongmei Liu, Alessandro Datti, Garnet Lau, Bryan W. Miller, Miriam Barrios-Rodiles
Jazyk: angličtina
Rok vydání: 2009
Předmět:
Systems biology
Regulator
Biology
Models
Biological
General Biochemistry
Genetics and Molecular Biology
Article
Cell Line
03 medical and health sciences
Mice
0302 clinical medicine
Axin Protein
RNA interference
Protein Interaction Mapping
Animals
Humans
030304 developmental biology
Adaptor Proteins
Signal Transducing
cancer biology
0303 health sciences
Gene knockdown
General Immunology and Microbiology
signal transduction
systems biology
Applied Mathematics
Calcium-Binding Proteins
Wnt signaling pathway
Signal transducing adaptor protein
Cell biology
High-Throughput Screening Assays
Repressor Proteins
Wnt Proteins
Computational Theory and Mathematics
030220 oncology & carcinogenesis
Ubiquitin-Conjugating Enzymes
RNA Interference
Signal transduction
General Agricultural and Biological Sciences
Carrier Proteins
Information Systems
Protein Binding
Zdroj: Molecular Systems Biology
Popis: 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 beta-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.
Databáze: OpenAIRE
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