Efflux Pump Control Alters Synthetic Gene Circuit Function

Junchen Diao, Daniel A. Charlebois, Dmitry Nevozhay, Zoltán Bódi, Csaba Pál, Gábor Balázsi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Synthetic biology aims to design new biological systems for predefined purposes, such as the controlled secretion of biofuels, pharmaceuticals, or other chemicals. Synthetic gene circuits regulating an efflux pump from the ATP-binding cassette (ABC) protein family could achieve this. However, ABC efflux pumps can also drive out intracellular inducer molecules that control the gene circuits. This will introduce an implicit feedback that could alter gene circuit function in ways that are poorly understood. Here, we used two synthetic gene circuits inducible by tetracycline family molecules to regulate the expression of a yeast ABC pump (Pdr5p) that pumps out the inducer. Pdr5p altered the dose-responses of the original gene circuits substantially in Saccharomyces cerevisiae. While one aspect of the change could be attributed to the efflux pumping function of Pdr5p, another aspect remained unexplained. Quantitative modeling indicated that reduced regulator gene expression in addition to efflux pump function could fully explain the altered dose-responses. These predictions were validated experimentally. Overall, we highlight how efflux pumps can alter gene circuit dynamics and demonstrate the utility of mathematical modeling in understanding synthetic gene circuit function in new circumstances.

Original languageEnglish
Pages (from-to)619-631
Number of pages13
JournalACS Synthetic Biology
Volume5
Issue number7
DOIs
Publication statusPublished - Jul 15 2016

Fingerprint

Synthetic Genes
Gene Regulatory Networks
Genes
Pumps
Networks (circuits)
Adenosinetriphosphate
Adenosine Triphosphate
Yeast
Synthetic Biology
Molecules
Biofuels
Biological systems
Regulator Genes
Tetracycline
Gene expression
Drug products
Saccharomyces cerevisiae
Carrier Proteins
Yeasts
Proteins

Keywords

  • efflux pump
  • feedback
  • stochastic simulation
  • synthetic gene circuit

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Biomedical Engineering

Cite this

Diao, J., Charlebois, D. A., Nevozhay, D., Bódi, Z., Pál, C., & Balázsi, G. (2016). Efflux Pump Control Alters Synthetic Gene Circuit Function. ACS Synthetic Biology, 5(7), 619-631. https://doi.org/10.1021/acssynbio.5b00154

Efflux Pump Control Alters Synthetic Gene Circuit Function. / Diao, Junchen; Charlebois, Daniel A.; Nevozhay, Dmitry; Bódi, Zoltán; Pál, Csaba; Balázsi, Gábor.

In: ACS Synthetic Biology, Vol. 5, No. 7, 15.07.2016, p. 619-631.

Research output: Contribution to journalArticle

Diao, J, Charlebois, DA, Nevozhay, D, Bódi, Z, Pál, C & Balázsi, G 2016, 'Efflux Pump Control Alters Synthetic Gene Circuit Function', ACS Synthetic Biology, vol. 5, no. 7, pp. 619-631. https://doi.org/10.1021/acssynbio.5b00154
Diao J, Charlebois DA, Nevozhay D, Bódi Z, Pál C, Balázsi G. Efflux Pump Control Alters Synthetic Gene Circuit Function. ACS Synthetic Biology. 2016 Jul 15;5(7):619-631. https://doi.org/10.1021/acssynbio.5b00154
Diao, Junchen ; Charlebois, Daniel A. ; Nevozhay, Dmitry ; Bódi, Zoltán ; Pál, Csaba ; Balázsi, Gábor. / Efflux Pump Control Alters Synthetic Gene Circuit Function. In: ACS Synthetic Biology. 2016 ; Vol. 5, No. 7. pp. 619-631.
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