Engineering dynamical control of cell fate switching using synthetic phospho-regulons.
Many cells can sense and respond to time-varying stimuli, selectively triggering changes in cell fate only in response to inputs of a particular duration or frequency. A common motif in dynamically controlled cells is a dual-timescale regulatory network: although long-term fate decisions are ultimately controlled by a slow-timescale switch (e.g., gene expression), input signals are first processed by a fast-timescale signaling layer, which is hypothesized to filter what dynamic information is efficiently relayed downstream. Directly testing the design principles of how dual-timescale circuits control dynamic sensing, however, has been challenging, because most synthetic biology methods have focused solely on rewiring transcriptional circuits, which operate at a single slow timescale. Here, we report the development of a modular approach for flexibly engineering phosphorylation circuits using designed phospho-regulon motifs. By then linking rapid phospho-feedback with slower downstream transcription-based bistable switches, we can construct synthetic dual-timescale circuits in yeast in which the triggering dynamics and the end-state properties of the ON state can be selectively tuned. These phospho-regulon tools thus open up the possibility to engineer cells with customized dynamical control.
Howard Hughes Medical Institute, San Francisco, CA 94158.|Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158.
Russell M
RM
Gordley
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158.|Graduate Group in Biophysics, University of California, San Francisco, CA 94158.
Reid E
RE
Williams
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158.|Graduate Group in Biophysics, University of California, San Francisco, CA 94158.
Caleb J
CJ
Bashor
Howard Hughes Medical Institute, San Francisco, CA 94158.
Jared E
JE
Toettcher
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158.
Shude
S
Yan
Howard Hughes Medical Institute, San Francisco, CA 94158; wendell.lim@ucsf.edu.|Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158.
Wendell A
WA
Lim
D060846:Cell Engineering; D019070:Cell Lineage; D053263:Gene Regulatory Networks; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010766:Phosphorylation; D018085:Regulon / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D058615:Synthetic Biology; D013997:Time Factors; D014158:Transcription, Genetic
D029701:Saccharomyces cerevisiae Proteins; C075580:FUS3 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases
dynamical control; phosphorylation; synthetic biology