title: Evolution of synthetic signaling scaffolds by recombination of modular protein domains.
abstract: Signaling scaffolds are proteins that interact via modular domains with multiple partners, regulating signaling networks in space and time and providing an ideal platform from which to alter signaling functions. However, to better exploit scaffolds for signaling engineering, it is necessary to understand the full extent of their modularity. We used a directed evolution approach to identify, from a large library of randomly shuffled protein interaction domains, variants capable of rescuing the signaling defect of a yeast strain in which Ste5, the scaffold in the mating pathway, had been deleted. After a single round of selection, we identified multiple synthetic scaffold variants with diverse domain architectures, able to mediate mating pathway activation in a pheromone-dependent manner. The facility with which this signaling network accommodates changes in scaffold architecture suggests that the mating signaling complex does not possess a single, precisely defined geometry into which the scaffold has to fit. These relaxed geometric constraints may facilitate the evolution of signaling networks, as well as their engineering for applications in synthetic biology.
authors.affiliation: Department of Cell and Systems Biology University of Toronto 25 Harbord Street, Toronto, Ontario M5S 3G5, Canada.
authors.forename: Andicus
authors.initials: A
authors.lastname: Lai
authors.affiliation: Department of Cell and Systems Biology University of Toronto 25 Harbord Street, Toronto, Ontario M5S 3G5, Canada.
authors.forename: Paloma M
authors.initials: PM
authors.lastname: Sato
authors.affiliation: Department of Cell and Systems Biology University of Toronto 25 Harbord Street, Toronto, Ontario M5S 3G5, Canada.
authors.forename: Sergio G
authors.initials: SG
authors.lastname: Peisajovich
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D016297:Mutagenesis, Site-Directed; D010675:Pheromones / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D010675:Pheromones; C080321:STE5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins
keywords: domain shuffling; evolution; scaffold; signaling networks; synthetic biology