Using engineered scaffold interactions to reshape MAP kinase pathway signaling dynamics.
Scaffold proteins link signaling molecules into linear pathways by physically assembling them into complexes. Scaffolds may also have a higher-order role as signal-processing hubs, serving as the target of feedback loops that optimize signaling amplitude and timing. We demonstrate that the Ste5 scaffold protein can be used as a platform to systematically reshape output of the yeast mating MAP kinase pathway. We constructed synthetic positive- and negative-feedback loops by dynamically regulating recruitment of pathway modulators to an artificial binding site on Ste5. These engineered circuits yielded diverse behaviors: ultrasensitive dose response, accelerated or delayed response times, and tunable adaptation. Protein scaffolds provide a flexible platform for reprogramming cellular responses and could be exploited to engineer cells with novel therapeutic and biotechnological functions.
Bashor
Caleb J
CJ
Department of Cellular and Molecular Pharmacology, University of California at San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
Helman
Noah C
NC
Yan
Shude
S
Lim
Wendell A
WA
D000222:Adaptation, Physiological; D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D025461:Feedback, Physiological; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D016350:Leucine Zippers; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D020935:MAP Kinase Signaling System; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000378:metabolism / Q000494:pharmacology; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D049490:Systems Biology / Q000379:methods
D048868:Adaptor Proteins, Signal Transducing; D047908:Intracellular Signaling Peptides and Proteins; C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C080321:STE5 protein, S cerevisiae; C079506:STE50 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C075580:FUS3 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases; C067018:Ste11 protein, S cerevisiae; C085791:MSG5 protein, S cerevisiae; D017027:Protein Tyrosine Phosphatases