Synthetic Gene Circuit-Mediated Monitoring of Endogenous Metabolites: Identification of GAL11 as a Novel Multicopy Enhancer of S‑Adenosylmethionine Level in Yeast

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 t...

Full description

Saved in:
Bibliographic Details
Published in:ACS synthetic biology 2013-08, Vol.2 (8), p.425-430
Main Authors: Umeyama, Taichi, Okada, Satoshi, Ito, Takashi
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Cloning, Molecular - methods
DNA Copy Number Variations - genetics
Enhancer Elements, Genetic - genetics
Gene Expression Regulation, Bacterial - genetics
Genes, Reporter - genetics
Genes, Synthetic - genetics
Mediator Complex - genetics
Metabolome - physiology
Monitoring, Physiologic - methods
S-Adenosylmethionine - analysis
S-Adenosylmethionine - genetics
S-Adenosylmethionine - metabolism
Saccharomyces cerevisiae - physiology
Saccharomyces cerevisiae Proteins - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:2161-5063
2161-5063
DOI:10.1021/sb300115n