Programming mRNA decay to modulate synthetic circuit resource allocation
| Autor: | Leanne Jade G. Chan, Ophelia S. Venturelli, Stefan Bauer, Mika Tei, Christopher J. Petzold, Adam P. Arkin |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Proteomics Computer science RNA Stability Messenger General Physics and Astronomy Proportional control Bioinformatics Transcriptome Synthetic biology Theoretical Models Translation factor Regulatory Elements Transcriptional Peptide sequence Feedback Physiological 0303 health sciences Multidisciplinary Escherichia coli Proteins DNA-Binding Proteins Metabolic Engineering Transcriptional Metabolic Networks and Pathways Biotechnology Science Circuit design Physiological 030106 microbiology Endoribonuclease Computational biology Biology General Biochemistry Genetics and Molecular Biology Feedback Metabolic engineering 03 medical and health sciences Endoribonucleases Genetics Escherichia coli Amino Acid Sequence RNA Messenger Gene 030304 developmental biology 030306 microbiology General Chemistry Feedback loop Models Theoretical Regulatory Elements 030104 developmental biology Resource allocation RNA Flux (metabolism) |
| Zdroj: | Nature communications, vol 8, iss 1 Venturelli, OS; Tei, M; Bauer, S; Chan, LJG; Petzold, CJ; & Arkin, AP. (2017). Programming mRNA decay to modulate synthetic circuit resource allocation. Nature Communications, 8. doi: 10.1038/ncomms15128. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/02v2x56n Nature Communications, Vol 8, Iss 1, Pp 1-11 (2017) |
| ISSN: | 2041-1723 |
| Popis: | Synthetic biomolecular networks embedded in host-cells compete with cellular processes for limited intracellular resources. Resource utilization is a major variable that impacts synthetic circuit behavior. Here we show that intracellular resources could be diverted from cellular operations to a synthetic circuit by programming global mRNA decay using the sequence-dependent endoribonuclease MazF. Synthetic circuit genes were protected from MazF activity by recoding the gene sequence to eliminate recognition sites, while preserving the amino acid sequence. The expression of a protected fluorescent reporter and the metabolic flux of a high-value metabolite gluconate were significantly enhanced using this genome-scale control strategy. Proteomics measurements discovered a key host translation factor in need of protection to optimize the resource redistribution activity. A dynamic computational model demonstrated that the MazF mRNA-decay feedback loop achieved proportional control of MazF levels in an optimal operating regime. RNA-seq time-series measurements of MazF-induced cells elucidated the dynamic shifts in the transcript abundance and discovered regulatory design elements that could be used to expand the control mechanisms of the MazF resource allocator. Together, these results demonstrated that manipulation of resource allocation is a tunable parameter that can be used to redirect resources away from cellular processes to synthetic circuits to enhance target functions. |
| Databáze: | OpenAIRE |
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