Systematic Interactome Mapping and Genetic Perturbation Analysis of a C. elegans TGF-β Signaling Network

• Published in: Molecular cell Vol. 13; no. 4; pp. 469 - 482
• Main Authors: Tewari, Muneesh, Hu, Patrick J., Ahn, Jin Sook, Ayivi-Guedehoussou, Nono, Vidalain, Pierre-Olivier, Li, Siming, Milstein, Stuart, Armstrong, Chris M., Boxem, Mike, Butler, Maurice D., Busiguina, Svetlana, Rual, Jean-François, Ibarrola, Nieves, Chaklos, Sabrina T., Bertin, Nicolas, Vaglio, Philippe, Edgley, Mark L., King, Kevin V., Albert, Patrice S., Vandenhaute, Jean, Pandey, Akhilesh, Riddle, Donald L., Ruvkun, Gary, Vidal, Marc
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 27.02.2004
• Subjects: Animals; Caenorhabditis elegans; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Cell Line; Chromosome Mapping; Cosmids; Gene Expression Regulation, Developmental; Genes, Helminth; Genomics; Humans; Kidney; Models, Biological; Mutation; Open Reading Frames; Proteome - metabolism; RNA, Small Interfering - metabolism; Signal Transduction; Transfection; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - metabolism; Transgenes; Two-Hybrid System Techniques
• ISSN: 1097-2765; 1097-4164
• DOI: 10.1016/S1097-2765(04)00033-4

To initiate a system-level analysis of C. elegans DAF-7/TGF-β signaling, we combined interactome mapping with single and double genetic perturbations. Yeast two-hybrid (Y2H) screens starting with known DAF-7/TGF-β pathway components defined a network of 71 interactions among 59 proteins. Coaffinity purification (co-AP) assays in mammalian cells confirmed the overall quality of this network. Systematic perturbations of the network using RNAi, both in wild-type and daf-7/TGF-β pathway mutant animals, identified nine DAF-7/TGF-β signaling modifiers, seven of which are conserved in humans. We show that one of these has functional homology to human SNO/SKI oncoproteins and that mutations at the corresponding genetic locus daf-5 confer defects in DAF-7/TGF-β signaling. Our results reveal substantial molecular complexity in DAF-7/TGF-β signal transduction. Integrating interactome maps with systematic genetic perturbations may be useful for developing a systems biology approach to this and other signaling modules.