title: Synthetic gene circuit-mediated monitoring of endogenous metabolites: identification of GAL11 as a novel multicopy enhancer of s-adenosylmethionine level in yeast.
abstract: Monitoring levels of key metabolites in living cells comprises a critical step in various investigations. The simplest approach to this goal is a fluorescent reporter gene using an endogenous promoter responsive to the metabolite. However, such a promoter is often not identified or even present in the species of interest. An alternative can be a synthetic gene circuit based on a heterologous pair consisting of a promoter and a transcription factor known to respond to the metabolite. We exploited the met operator and MetJ repressor of Escherichia coli, the interaction between which depends on S-adenosylmethionine (SAM), to construct synthetic gene circuits that report SAM levels in Saccharomyces cerevisiae. Using a dual-input circuit that outputs selection marker genes in a doxycycline-tunable manner, we screened a genomic library to identify GAL11 as a novel multicopy enhancer of SAM levels. These results demonstrate the potential and utility of synthetic gene circuit-mediated metabolite monitoring.
authors: Taichi
authors: T
authors: Umeyama
authors: Satoshi
authors: S
authors: Okada
authors: Takashi
authors: T
authors: Ito
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D056915:DNA Copy Number Variations / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics*; D005813:Genes, Synthetic / Q000235:genetics*; D056892:Mediator Complex / Q000235:genetics*; D055442:Metabolome / Q000502:physiology*; D008991:Monitoring, Physiologic / Q000379:methods; D012436:S-Adenosylmethionine / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; C058337:GAL11 protein, S cerevisiae; D056892:Mediator Complex; D029701:Saccharomyces cerevisiae Proteins; D012436:S-Adenosylmethionine