A scalable framework for the discovery of functional helicase substrates and helicase-driven regulatory switches
| Autor: | Mildred Delaleau, Eric Eveno, Isabelle Simon, Annie Schwartz, Marc Boudvillain |
|---|---|
| Přispěvatelé: | Boudvillain, Marc, Toward an Integrated View of Transcription Terminator Rho Functions and Mechanisms - - IntegRhoMe2015 - ANR-15-CE11-0024 - AAPG2015 - VALID, Exploiter l'activité ARN hélicase à des fins de (ribo)régulation synthétique - - HELISWITCH2019 - ANR-19-CE44-0009 - AAPG2019 - VALID, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), ANR-15-CE11-0024,IntegRhoMe,Toward an Integrated View of Transcription Terminator Rho Functions and Mechanisms(2015), ANR-19-CE44-0009,HELISWITCH,Exploiter l'activité ARN hélicase à des fins de (ribo)régulation synthétique(2019) |
| Rok vydání: | 2022 |
| Předmět: |
riboswitches
Binding Sites Multidisciplinary [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] helicases SELEX Aptamer Technique DNA Helicases [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology Ligands Substrate Specificity Rho Riboswitch Transcription Termination Genetic Escherichia coli [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] transcription termination |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, 2022, 119 (38), ⟨10.1073/pnas.2209608119⟩ |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | Helicases are ubiquitous motor enzymes that remodel nucleic acids (NA) and NA–protein complexes in key cellular processes. To explore the functional repertoire and specificity landscape of helicases, we devised a screening scheme—Helicase-SELEX (Systematic Evolution of Ligands by EXponential enrichment)—that enzymatically probes substrate and cofactor requirements at global scale. Using the transcription termination Rho helicase of Escherichia coli as a prototype for Helicase-SELEX, we generated a genome-wide map of Rho utilization ( Rut ) sites. The map reveals many features, including promoter- and intrinsic terminator-associated Rut sites, bidirectional Rut tandems, and cofactor-dependent Rut sites with inverted G > C skewed compositions. We also implemented an H-SELEX variant where we used a model ligand, serotonin, to evolve synthetic Rut sites operating in vitro and in vivo in a ligand-dependent manner. Altogether, our data illustrate the power and flexibility of Helicase-SELEX to seek constitutive or conditional helicase substrates in natural or synthetic NA libraries for fundamental or synthetic biology discovery. |
| Databáze: | OpenAIRE |
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