Abstract
Phenotypic variations of an organism may arise from alterations of cellular networks, ranging from the complete loss of a gene product to the specific perturbation of a single molecular interaction. In interactome networks that are modeled as nodes (macromolecules) connected by edges (interactions), these alterations can be thought of as node removal and edge-specific or “edgetic” perturbations, respectively. Here we present two complementary strategies, forward and reverse edgetics, to investigate the phenotypic outcomes of edgetic perturbations of binary protein–protein interaction networks. Both approaches are based on the yeast two-hybrid system (Y2H). The first allows the determination of the interaction profile of proteins encoded by alleles with known phenotypes to identify edgetic alleles. The second is used to directly isolate edgetic alleles for subsequent in vivo characterization.
B. Charloteaux, Q. Zhong, and M. Dreze contributed equally to this work.
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Acknowledgements
Special thanks to R. Vidal for her help with the manuscript. This work was supported by the Ellison Foundation and the WM Keck Foundation (MV); NIH grants R01-HG001715 from NHGRI and NIGMS (MV and DEH), U01-CA105423 (PI, S Orkin, project leader, MV), and U54-CA112952 (PI, J Nevins, subcontract, MV) and R33 CA105405, R33 CA132073, and R21/R33 CA081658 (MV) from NCI, and by Institute Sponsored Research funds from the Dana-Farber Cancer Institute Strategic Initiative awarded to CCSB. M.V. is a “Chercheur Qualifié Honoraire” from 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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© 2011 Humana Press
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Charloteaux, B., Zhong, Q., Dreze, M., Cusick, M.E., Hill, D.E., Vidal, M. (2011). Protein–Protein Interactions and Networks: Forward and Reverse Edgetics. In: Castrillo, J., Oliver, S. (eds) Yeast Systems Biology. Methods in Molecular Biology, vol 759. Humana Press. https://doi.org/10.1007/978-1-61779-173-4_12
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DOI: https://doi.org/10.1007/978-1-61779-173-4_12
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