Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120

Aims: The ferric uptake regulator (Fur) is the main transcriptional regulator of genes involved in iron homeostasis in most prokaryotes. FurA from Anabaena sp. PCC 7120 contains five cysteine residues, four of them arranged in two redox-active CXXC motifs. The protein needs not only metal but also r...

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Autores: Botello-Morte, L, Pellicer, S, Sein-Echaluce, VC, Contreras, LM, Neira, JL, Abian, O, Velazquez-Campoy, A, Peleato, ML, Fillat, MF, Bes, MT
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Institución:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
Repositorio:r-FISABIO. Repositorio Institucional de Producción Científica
OAI Identifier:oai:fisabio.fundanetsuite.com:p12839
Acceso en línea:https://fisabio.portalinvestigacion.com/publicaciones/12839
Access Level:acceso abierto
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spelling Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120Botello-Morte, LPellicer, SSein-Echaluce, VCContreras, LMNeira, JLAbian, OVelazquez-Campoy, APeleato, MLFillat, MFBes, MTAims: The ferric uptake regulator (Fur) is the main transcriptional regulator of genes involved in iron homeostasis in most prokaryotes. FurA from Anabaena sp. PCC 7120 contains five cysteine residues, four of them arranged in two redox-active CXXC motifs. The protein needs not only metal but also reducing conditions to remain fully active in vitro. Through a mutational study of the cysteine residues present in FurA, we have investigated their involvement in metal and DNA binding. Results: Residue C-101 that belongs to a conserved CXXC motif plays an essential role in both metal and DNA binding activities in vitro. Substitution of C-101 by serine impairs DNA and metal binding abilities of FurA. Isothermal titration calorimetry measurements show that the redox state of C-101 is responsible for the protein ability to coordinate the metal corepressor. Moreover, the redox state of C-101 varies with the presence or absence of C-104 or C-133, suggesting that the environments of these cysteines are mutually interdependent. Innovation: We propose that C-101 is part of a thiol/disulfide redox switch that determines FurA ability to bind the metal corepressor. Conclusion: This mechanism supports a novel feature of a Fur protein that emerges as a regulator, which connects the response to changes in the intracellular redox state and iron management in cyanobacteria.MARY ANN LIEBERT, INC2016info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://fisabio.portalinvestigacion.com/publicaciones/12839ANTIOXIDANTS & REDOX SIGNALINGISSN: 15230864ISSNe: 15577716reponame:r-FISABIO. Repositorio Institucional de Producción Científicainstname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)Inglésinfo:eu-repo/semantics/openAccessoai:fisabio.fundanetsuite.com:p128392026-06-11T12:45:17Z
dc.title.none.fl_str_mv Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
title Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
spellingShingle Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
Botello-Morte, L
title_short Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
title_full Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
title_fullStr Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
title_full_unstemmed Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
title_sort Cysteine Mutational Studies Provide Insight into a Thiol-Based Redox Switch Mechanism of Metal and DNA Binding in FurA from Anabaena sp PCC 7120
dc.creator.none.fl_str_mv Botello-Morte, L
Pellicer, S
Sein-Echaluce, VC
Contreras, LM
Neira, JL
Abian, O
Velazquez-Campoy, A
Peleato, ML
Fillat, MF
Bes, MT
author Botello-Morte, L
author_facet Botello-Morte, L
Pellicer, S
Sein-Echaluce, VC
Contreras, LM
Neira, JL
Abian, O
Velazquez-Campoy, A
Peleato, ML
Fillat, MF
Bes, MT
author_role author
author2 Pellicer, S
Sein-Echaluce, VC
Contreras, LM
Neira, JL
Abian, O
Velazquez-Campoy, A
Peleato, ML
Fillat, MF
Bes, MT
author2_role author
author
author
author
author
author
author
author
author
description Aims: The ferric uptake regulator (Fur) is the main transcriptional regulator of genes involved in iron homeostasis in most prokaryotes. FurA from Anabaena sp. PCC 7120 contains five cysteine residues, four of them arranged in two redox-active CXXC motifs. The protein needs not only metal but also reducing conditions to remain fully active in vitro. Through a mutational study of the cysteine residues present in FurA, we have investigated their involvement in metal and DNA binding. Results: Residue C-101 that belongs to a conserved CXXC motif plays an essential role in both metal and DNA binding activities in vitro. Substitution of C-101 by serine impairs DNA and metal binding abilities of FurA. Isothermal titration calorimetry measurements show that the redox state of C-101 is responsible for the protein ability to coordinate the metal corepressor. Moreover, the redox state of C-101 varies with the presence or absence of C-104 or C-133, suggesting that the environments of these cysteines are mutually interdependent. Innovation: We propose that C-101 is part of a thiol/disulfide redox switch that determines FurA ability to bind the metal corepressor. Conclusion: This mechanism supports a novel feature of a Fur protein that emerges as a regulator, which connects the response to changes in the intracellular redox state and iron management in cyanobacteria.
publishDate 2016
dc.date.none.fl_str_mv 2016
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://fisabio.portalinvestigacion.com/publicaciones/12839
url https://fisabio.portalinvestigacion.com/publicaciones/12839
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MARY ANN LIEBERT, INC
publisher.none.fl_str_mv MARY ANN LIEBERT, INC
dc.source.none.fl_str_mv ANTIOXIDANTS & REDOX SIGNALING
ISSN: 15230864
ISSNe: 15577716
reponame:r-FISABIO. Repositorio Institucional de Producción Científica
instname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
instname_str Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
reponame_str r-FISABIO. Repositorio Institucional de Producción Científica
collection r-FISABIO. Repositorio Institucional de Producción Científica
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.812429

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