Increasing the Scalability of Toxin-Intein Orthogonal Combinations

Inteins are proteins embedded into host proteins from which they are excised in an autocatalytic reaction. Specifically, split inteins are separated into two independent fragments that reconstitute the host protein during the catalytic process. We recently developed a novel strategy for the specific...

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Detalles Bibliográficos
Autores: López-Igual, Rocío, Dorado-Morales, Pedro, Mazel, Didier
Tipo de recurso: artículo
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/354660
Acceso en línea:http://hdl.handle.net/10261/354660
https://api.elsevier.com/content/abstract/scopus_id/85147145110
Access Level:acceso abierto
Palabra clave:Bacterial killing
Inteins
Microbial synthetic biology
Protein splicing
Toxin−antitoxin systems
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spelling Increasing the Scalability of Toxin-Intein Orthogonal CombinationsLópez-Igual, RocíoDorado-Morales, PedroMazel, DidierBacterial killingInteinsMicrobial synthetic biologyProtein splicingToxin−antitoxin systemsInteins are proteins embedded into host proteins from which they are excised in an autocatalytic reaction. Specifically, split inteins are separated into two independent fragments that reconstitute the host protein during the catalytic process. We recently developed a novel strategy for the specific killing of pathogenic and antibiotic resistant bacteria based on toxin-intein combinations. Bacterial type II toxin-antitoxin systems are protein modules in which the toxin can provoke cell death whereas the antitoxin inhibits toxin activity. Although our previous system was based on a split intein (iDnaE) and the CcdB toxin, we demonstrated that iDnaE is able to reconstitute four different toxins. To expand the applicability of our system by widening the repertoire of toxin-intein combinations for complex set-ups, we introduced a second intein, iDnaX, which was artificially split. We demonstrate that iDnaX is able to reconstitute the four toxins, and we manage to reduce its scar size to facilitate their use. In addition, we prove the orthogonality of both inteins (iDnaE and iDnaX) through a toxin reconstitution assay, thus opening the possibility for complex set-ups based on these toxin-intein modules. This could be used to develop specific antimicrobial and other biotechnological applications.This work was supported by the Institut Pasteur (to D.M.’s unit), the Centre National de la Recherche Scientifique (Grant No. CNRS-UMR 3525) (to D.M.), PLASWIRES 612146/FP7-FET-Proactive (to D.M.’s unit, and R.L.-I.), the French Government’s Investissement d’Avenir program, Laboratoire d’Excellence “Integrative Biology of Emerging Infectious Diseases” (Grant No. ANR-10-LABX-62-IBEID to D.M.’s unit), the Fondation pour la Recherche Médicale (Grant No. EQU202103012569 to P.D.-M. to D.M.), and Grant PID2019-104784RJ-I00 MCIN/AEI/10.13039/501100011033 Spain and Grant BFU2017-88202-P MCIN/AEI/10.13039/501100011033 Spain, cofinanced by ERDF A Way of Making Europe to R.L.-I.Peer reviewedAmerican Chemical SocietyInstitut PasteurCentre National de la Recherche Scientifique (France)Fondation pour la Recherche MédicaleMinisterio de Ciencia, Innovación y Universidades (España)López-Igual, Rocío [0000-0002-2369-1583]Dorado-Morales, Pedro [0000-0001-5760-1999]Mazel, Didier [0000-0001-6482-6002]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501application/pdfhttp://hdl.handle.net/10261/354660https://api.elsevier.com/content/abstract/scopus_id/85147145110reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104784RJ-I00https://doi.org/10.1021/acssynbio.2c00477Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3546602026-05-22T06:33:51Z
dc.title.none.fl_str_mv Increasing the Scalability of Toxin-Intein Orthogonal Combinations
title Increasing the Scalability of Toxin-Intein Orthogonal Combinations
spellingShingle Increasing the Scalability of Toxin-Intein Orthogonal Combinations
López-Igual, Rocío
Bacterial killing
Inteins
Microbial synthetic biology
Protein splicing
Toxin−antitoxin systems
title_short Increasing the Scalability of Toxin-Intein Orthogonal Combinations
title_full Increasing the Scalability of Toxin-Intein Orthogonal Combinations
title_fullStr Increasing the Scalability of Toxin-Intein Orthogonal Combinations
title_full_unstemmed Increasing the Scalability of Toxin-Intein Orthogonal Combinations
title_sort Increasing the Scalability of Toxin-Intein Orthogonal Combinations
dc.creator.none.fl_str_mv López-Igual, Rocío
Dorado-Morales, Pedro
Mazel, Didier
author López-Igual, Rocío
author_facet López-Igual, Rocío
Dorado-Morales, Pedro
Mazel, Didier
author_role author
author2 Dorado-Morales, Pedro
Mazel, Didier
author2_role author
author
dc.contributor.none.fl_str_mv Institut Pasteur
Centre National de la Recherche Scientifique (France)
Fondation pour la Recherche Médicale
Ministerio de Ciencia, Innovación y Universidades (España)
López-Igual, Rocío [0000-0002-2369-1583]
Dorado-Morales, Pedro [0000-0001-5760-1999]
Mazel, Didier [0000-0001-6482-6002]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Bacterial killing
Inteins
Microbial synthetic biology
Protein splicing
Toxin−antitoxin systems
topic Bacterial killing
Inteins
Microbial synthetic biology
Protein splicing
Toxin−antitoxin systems
description Inteins are proteins embedded into host proteins from which they are excised in an autocatalytic reaction. Specifically, split inteins are separated into two independent fragments that reconstitute the host protein during the catalytic process. We recently developed a novel strategy for the specific killing of pathogenic and antibiotic resistant bacteria based on toxin-intein combinations. Bacterial type II toxin-antitoxin systems are protein modules in which the toxin can provoke cell death whereas the antitoxin inhibits toxin activity. Although our previous system was based on a split intein (iDnaE) and the CcdB toxin, we demonstrated that iDnaE is able to reconstitute four different toxins. To expand the applicability of our system by widening the repertoire of toxin-intein combinations for complex set-ups, we introduced a second intein, iDnaX, which was artificially split. We demonstrate that iDnaX is able to reconstitute the four toxins, and we manage to reduce its scar size to facilitate their use. In addition, we prove the orthogonality of both inteins (iDnaE and iDnaX) through a toxin reconstitution assay, thus opening the possibility for complex set-ups based on these toxin-intein modules. This could be used to develop specific antimicrobial and other biotechnological applications.
publishDate 2023
dc.date.none.fl_str_mv 2023
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/354660
https://api.elsevier.com/content/abstract/scopus_id/85147145110
url http://hdl.handle.net/10261/354660
https://api.elsevier.com/content/abstract/scopus_id/85147145110
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104784RJ-I00
https://doi.org/10.1021/acssynbio.2c00477
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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