Fig. 1

Language component acquisition: genome-mining yields a scalable pool of peptide-GPCR interfaces. a Pipeline for component harvest and communication assembly. Upper panel: mining of Ascomycete genomes yields a scalable pool of peptide-GPCR pairs. Middle panel: GPCR activation can be coupled to peptide secretion to establish two-cell communication links. Each cell senses an incoming peptide signal via a specific GPCR, with GPCR activation leading to secretion of an orthogonal user-chosen peptide. The secreted peptide serves as the outgoing signal sensed by the second cell. Lower panel: Scalable communication networks can be assembled in a plug-and play manner using the two-cell communication links. b GPCRs and peptides can be swapped by simple DNA cloning. Conservation in both GPCR signal transduction and peptide secretion enables scalable communication without any additional strain engineering. Mating GPCRs couple to the S. cerevisiae Gα protein (Gpa1) and signals are transduced through a MAP-kinase-mediated phosphorylation cascade. Gene activation is then mediated by the transcription factor Ste12 through binding of a pheromone response element (PRE, gray) in the promoters of mating-associated genes (e.g., FUS1 and FIG1, used herein to control synthetic constructs of choice). Peptides are translated by the ribosome as pre-pro peptides. Pre-pro peptide architecture is conserved and starts with an N-terminal secretion signal (light blue), followed by Ste13 and Kex2 recognition sites (gray and yellow, respectively). Mature secreted peptides (red) are processed while trafficking through the ER and Golgi. The conserved pre-pro-peptide architecture enables the bioinformatic deorphanization of fungal GPCRs by inference of mature peptide sequences from precursor genes. c Most genome-mined peptide-GPCR pairs are functional in yeast. Functionality of 45 peptide-GPCR pairs was evaluated by on/off testing using 40 μM cognate peptide and fluorescence as read-out. GPCRs are organized by percent amino acid identity to the Sc.Ste2. Non-functional GPCRs (those that give a signal difference < 3 standard deviations) are highlighted in red; constitutive GPCRs are highlighted in green. GPCR nomenclature corresponds to species names (Supplementary Table 1). Experiments were performed in triplicate and full data sets with errors (standard deviation) and individual data points are given in Supplementary Figure 3