Design principles of dose-response alignment in coupled GTPase switches

"Dose-response alignment" (DoRA), where the downstream response of cellular signaling path-ways closely matches the fraction of activated receptor, can improve the fidelity of dose information transmission. It is believed that a key component for DoRA is negative feedback and thus a natural question that arises is whether there exist design principles for signaling motifs within such negative feedback loops, which may enable these motifs to attain near-perfect DoRA. Here, we investigated several model formulations of an experimentally validated circuit that couples two molecular switches--mGTPase (monomeric GTPase) and tGTPase (heterotrimeric GTPases) -- with negative feedback loops. We find that, in the absence of feedback, the low and intermediate mGTPase activation levels benefit DoRA in the mass action and Hill-function models, respectively. In other cases, where the mass action model with a high mGTPase activation level or the Hill-function model with a non-intermediate mGTPase activation level, the DoRA can be improved by adding negative feedback loops. Furthermore, we found that DoRA in a longer cascade (i.e., tGTPase) can be obtained using Hill-function kinetics under certain conditions. In summary, we show how ranges of activity of mGTPase, reaction kinetics, the negative feedback, and the cascade length affect DoRA. This work provides a framework for improving the DoRA performance in signaling motifs with negative feedback loops. Significance StatementDose-response alignment helps cells faithfully transmit dose information; how this alignment is achieved in motifs with negative feedback is unclear. Through rigorous studies interrogating a naturally occurring motif comprised of two species of GTPases coupled by negative feedback loops, this work reveals the versatile roles of negative feedback loops and GTPase regulators on DoRA. We find that the negative feedback can enhance DoRA only with specific kinetic forms and with certain ranges of GTPases activation levels. This knowledge advances our understanding of the role of negative feedback on DoRA and sheds light on the importance of dynamic range of signaling processes as an essential determinant of how cells transfer information about stimuli. Findings can help design signaling circuits with better DoRA behavior, and ultimately augment cell signaling studies.