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dc.contributor.author
Tomatis, Pablo Emiliano
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dc.contributor.author
Schütz, Marco
dc.contributor.author
Umudumov, Elina
dc.contributor.author
Plückthun, Andreas
dc.date.available
2023-01-24T14:18:42Z
dc.date.issued
2019-12
dc.identifier.citation
Tomatis, Pablo Emiliano; Schütz, Marco; Umudumov, Elina; Plückthun, Andreas; Mutations in sigma 70 transcription factor improves expression of functional eukaryotic membrane proteins in Escherichia coli; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-14
dc.identifier.issn
2045-2322
dc.identifier.uri
http://hdl.handle.net/11336/185415
dc.description.abstract
Eukaryotic integral membrane proteins (IMPs) are difficult to study due to low functional expression levels. To investigate factors for efficient biogenesis of eukaryotic IMPs in the prokaryotic model organism Escherichia coli, important, e.g., for isotope-labeling for NMR, we selected for E. coli cells expressing high levels of functional G protein-coupled receptors (GPCRs) by FACS. Utilizing an E. coli strain library with all non-essential genes systematically deleted, we unexpectedly discovered upon whole-genome sequencing that the improved phenotype was not conferred by the deleted genes but by various subtle alterations in the “housekeeping” sigma 70 factor (RpoD). When analyzing effects of the rpoD mutations at the transcriptome level we found that toxic effects incurred on wild-type E. coli during receptor expression were diminished by two independent and synergistic effects: a slower but longer-lasting GPCR biosynthesis and an optimized transcriptional pattern, augmenting growth and expression at low temperature, setting the basis for further bacterial strain engineering.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Nature Publishing Group
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/
dc.subject
MEMBRANE PROTEINS
dc.subject
G PROTEIN-COUPLED RECEPTORS
dc.subject
EXPRESSION SYSTEMS
dc.subject
GENE EXPRESSION ANALYSIS
dc.subject
Bacterial secretion
dc.subject
Transcriptional Regulatory elements
dc.subject.classification
Bioquímica y Biología Molecular
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Mutations in sigma 70 transcription factor improves expression of functional eukaryotic membrane proteins in Escherichia coli
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2020-11-25T18:00:31Z
dc.journal.volume
9
dc.journal.number
1
dc.journal.pagination
1-14
dc.journal.pais
Reino Unido
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dc.description.fil
Fil: Tomatis, Pablo Emiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
dc.description.fil
Fil: Schütz, Marco. Universitat Zurich; Suiza
dc.description.fil
Fil: Umudumov, Elina. Universitat Zurich; Suiza
dc.description.fil
Fil: Plückthun, Andreas. Universitat Zurich; Suiza
dc.journal.title
Scientific Reports
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://doi.org/10.1038/s41598-019-39492-9
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