Cellular assays identify barriers impeding iron-sulfur enzyme activity in a non-native prokaryotic host
In: EISSN: 2050-084X ; eLife ; https://hal.science/hal-03634503 ; eLife, 2022, 11, pp.e70936. ⟨10.7554/eLife.70936⟩, 2022
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International audience ; Iron-sulfur (Fe-S) clusters are ancient and ubiquitous protein cofactors and play irreplaceable roles in many metabolic and regulatory processes. Fe-S clusters are built and distributed to Fe-S enzymes by dedicated protein networks. The core components of these networks are widely conserved and highly versatile. However, Fe-S proteins and enzymes are often inactive outside their native host species. We sought to systematically investigate the compatibility of Fe-S networks with non-native Fe-S enzymes. By using collections of Fe-S enzyme orthologs representative of the entire range of prokaryotic diversity, we uncovered a striking correlation between phylogenetic distance and probability of functional expression. Moreover, coexpression of a heterologous Fe-S biogenesis pathway increases the phylogenetic range of orthologs that can be supported by the foreign host. We also find that Fe-S enzymes that require specific electron carrier proteins are rarely functionally expressed unless their taxon-specific reducing partners are identified and co-expressed. We demonstrate how these principles can be applied to improve the activity of a radical S-adenosyl methionine(rSAM) enzyme from a Streptomyces antibiotic biosynthesis pathway in Escherichia coli. Our results clarify how oxygen sensitivity and incompatibilities with foreign Fe-S and electron transfer networks each impede heterologous activity. In particular, identifying compatible electron transfer proteins and heterologous Fe-S biogenesis pathways may prove essential for engineering functional Fe-S enzyme-dependent pathways.
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Cellular assays identify barriers impeding iron-sulfur enzyme activity in a non-native prokaryotic host
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Autor/in / Beteiligte Person: | D’angelo, Francesca ; Fernández-Fueyo, Elena ; Garcia, Pierre Simon ; Shomar, Héléna ; Pelosse, Martin ; Rebelo Manuel, Rita ; Büke, Ferhat ; Liu, Siyi ; van den Broek, Niels ; Duraffourg, Nicolas ; de Ram, Carol ; Pabst, Martin ; Bouveret, Emmanuelle ; Gribaldo, Simonetta ; Py, Béatrice ; Ollagnier de Choudens, Sandrine ; Barras, Frédéric ; Bokinsky, Grégory ; Adaptation au stress et Métabolisme chez les entérobactéries - Stress adaptation and metabolism in enterobacteria (SAMe) ; Université Paris Cité (UPCité)-Microbiologie Intégrative et Moléculaire (UMR6047) ; Institut Pasteur Paris (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur Paris (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) ; Kavli Institute of Nanosciences Delft (KI-NANO) ; Delft University of Technology (TU Delft) ; Biologie Évolutive de la Cellule Microbienne - Evolutionary Biology of the Microbial Cell ; (BIOCAT), Biocatalyse ; Laboratoire de Chimie et Biologie des Métaux (LCBM - UMR 5249) ; Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG) ; Direction de Recherche Fondamentale (CEA) (DRF (CEA)) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA) ; Laboratoire de chimie bactérienne (LCB) ; Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS) ; Catalyse Bioinorganique et Environnement (BioCE ) ; This project (IRONPLUGNPLAY) has received funding from the European Union’s Horizon 2020 research and innovation programme under grant 722361 and was supported by grants from Agence Nationale Recherche (ANR): ANR-10-LABX-62-IBEID, the LabEx ARCANE (ANR-11-LABX-0003-01), and the CBH-EUR-GS (ANR-17-EURE-0003). Synthetic DNA prepared by the Joint Genome Institute DNA Synthesis Program was provided to GB (awarded proposal 503636). The work conducted by the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User Facility, is supported under Contract No. DE-AC02-05CH11231. SL was supported by the China Scholarship Council. ; ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010) ; ANR-11-LABX-0003,ARCANE,Grenoble, une chimie bio-motivée(2011) ; ANR-17-EURE-0003,CBH-EUR-GS,CBH-EUR-GS(2017) ; European Project: 722361,ERA CoBioTech |
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Zeitschrift: | EISSN: 2050-084X ; eLife ; https://hal.science/hal-03634503 ; eLife, 2022, 11, pp.e70936. ⟨10.7554/eLife.70936⟩, 2022 |
Veröffentlichung: | HAL CCSD ; eLife Sciences Publication, 2022 |
Medientyp: | academicJournal |
DOI: | 10.7554/eLife.70936 |
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