An organism’s ability to sense, interpret, and respond to its environment is an essential requirement of life itself. At the cellular level, this process of connecting signals to behaviors is governed by a diverse set of mechanisms such as mitogen activated protein (MAP) kinase cascades, which are broadly conserved across animals, plants and fungi. Alterations to these connections that arise from evolution or disease can have significant phenotypic effects. MAP kinases transmit signals by colocalizing with a target protein and altering its structure through phosphorylation and thereby producing downstream behavioral changes in the cell. Engineering novel phosphorylation triggered interaction domains will allow MAP kinase signaling networks to be rewired to correct pathological dysfunction or to generate novel responses to environmental signals. We intend to use an engineered protein scaffold with a phosphorylatable motif and an occluded binding motif, which was designed by collaborators in the Baker lab, to create a phosphorylation triggered interaction motif in Saccharomyces cerevisiae. By rewiring the S. cerevisiae mating MAP kinase cascade to phosphorylate this scaffold and incorporating a yeast three hybrid output, we hope to demonstrate the function of this new class of interaction domains in vivo. We hypothesize that upon phosphorylation, the scaffold will undergo a conformational change revealing the previously occluded binding motif, allowing another protein to bind and produce downstream effects. Such novel interaction domains will serve as a “toolbox” to allow the rewiring of MAP kinase cascades in any context in which they naturally occur. This approach could be used to fix pathological miswiring of MAP kinase cascades that contribute to anomalies of development or diseases such as cancer. Additionally, the creation of new connections using these interaction domains could be utilized to engineer organisms or cells with useful functions such as plants with enhanced defensive traits or cancer targeting T-cells.