Genes and gene products do not function in isolation but within highly interconnected 'interactome' networks, modeled as graphs of nodes and edges representing macromolecules and interactions between them, respectively. We propose to investigate genotype-phenotype associations by methodical use of alleles that lack single interactions, while retaining all others, in contrast to genetic approaches designed to eliminate gene products completely. We describe an integrated strategy based on the reverse yeast two-hybrid system to isolate and characterize such edge-specific, or 'edgetic', alleles. We established a proof of concept with CED-9, a Caenorhabditis elegans BCL2 ortholog. Using ced-9 edgetic alleles, we uncovered a new potential functional link between apoptosis and a centrosomal protein. This approach is amenable to higher throughput and is particularly applicable to interactome network analysis in organisms for which transgenesis is straightforward.
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This paper is dedicated to the memory of Stan Korsmeyer. We thank the members of the Vidal Lab and of the Dana-Farber Cancer Institute CCSB and particularly A.-R. Carvunis for helpful discussions. This work was supported by US National Institutes of Health (NIH) grants R01 HG001715 from the National Human Genomics Research Institute (NHGRI) and National Institute of General Medical Sciences (NIGMS) and grants R33 CA105405, R33 CA132073 and R21/R33 CA081658 from the US National Cancer Institute (NCI) (M.V.), U01 CA105423 from the NCI (principal investigator, S. Orkin; project leader, M.V.) and by Institute Sponsored Research funds from the Dana-Farber Cancer Institute Strategic Initiative awarded to CCSB. M.D. and G.L. were supported by a 'Research Fellow' fellowship from the Fonds de la Recherche Scientifique (FRS-FNRS, French Community of Belgium). B.C. was supported by the Belgian Program on Interuniversity Attraction Poles initiated by the Federal Office for Scientific, Technical and Cultural Affairs (IAP P6/19 PROFUSA). S.M. was supported by an NIH National Research Service Award training grant fellowship (T32CA09361). P.-O.V. was supported by a European Molecular Biology Organization long-term fellowship 61-2002. Support was provided by the Leukemia Research Foundation to M.B. M.V. and R.B. thank the 'Fonds de la Recherche Scientifique (FRS-FNRS, French Community of Belgium)'. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NCI, NHGRI, NIGMS or the NIH.
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Dreze, M., Charloteaux, B., Milstein, S. et al. 'Edgetic' perturbation of a C. elegans BCL2 ortholog. Nat Methods 6, 843–849 (2009). https://doi.org/10.1038/nmeth.1394
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