Functional asymmetry and chemical reactivity of CsoR family persulfide sensors

Autores
Fakhoury, Joseph N; Zhang, Yifan; Edmonds, Katherine A; Bringas, Mauro; Luebke, Justin L; Gonzalez Gutierrez, Giovanni; Capdevila, Daiana Andrea; Giedroc, David Peter
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
CstR is a persulfide-sensing member of the functionally diverse copper-sensitive operon repressor (CsoR) superfamily. While CstR regulates the bacterial response to hydrogen sulfide (H2S) and more oxidized reactive sulfur species (RSS) in Gram-positive pathogens, other dithiol-containing CsoR proteins respond to host derived Cu(I) toxicity, sometimes in the same bacterial cytoplasm, but without regulatory crosstalk in cells. It is not clear what prevents this crosstalk, nor the extent to which RSS sensors exhibit specificity over other oxidants. Here, we report a sequence similarity network (SSN) analysis of the entire CsoR superfamily, which together with the first crystallographic structure of a CstR and comprehensive mass spectrometry-based kinetic profiling experiments, reveal new insights into the molecular basis of RSS specificity in CstRs. We find that the more N-terminal cysteine is the attacking Cys in CstR and is far more nucleophilic than in a CsoR. Moreover, our CstR crystal structure is markedly asymmetric and chemical reactivity experiments reveal the functional impact of this asymmetry. Substitution of the Asn wedge between the resolving and the attacking thiol with Ala significantly decreases asymmetry in the crystal structure and markedly impacts the distribution of species, despite adopting the same global structure as the parent repressor. Companion NMR, SAXS and molecular dynamics simulations reveal that the structural and functional asymmetry can be traced to fast internal dynamics of the tetramer. Furthermore, this asymmetry is preserved in all CstRs and with all oxidants tested, giving rise to markedly distinct distributions of crosslinked products. Our exploration of the sequence, structural, and kinetic features that determine oxidant-specificity suggest that the product distribution upon RSS exposure is determined by internal flexibility.
Fil: Fakhoury, Joseph N. Indiana University; Estados Unidos
Fil: Zhang, Yifan. Indiana University; Estados Unidos
Fil: Edmonds, Katherine A. Indiana University; Estados Unidos
Fil: Bringas, Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Luebke, Justin L. Indiana University; Estados Unidos
Fil: Gonzalez Gutierrez, Giovanni. Indiana University; Estados Unidos
Fil: Capdevila, Daiana Andrea. Indiana University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Giedroc, David Peter. Indiana University; Estados Unidos
Materia
TRANSCRIPTIONAL REGULATORS
RSS
STREPTOCOCCUS PNEUMONIAE
FUNCTIONAL ASSYMMMETRY
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/167149
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/167149
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Functional asymmetry and chemical reactivity of CsoR family persulfide sensorsFakhoury, Joseph NZhang, YifanEdmonds, Katherine ABringas, MauroLuebke, Justin LGonzalez Gutierrez, GiovanniCapdevila, Daiana AndreaGiedroc, David PeterTRANSCRIPTIONAL REGULATORSRSSSTREPTOCOCCUS PNEUMONIAEFUNCTIONAL ASSYMMMETRYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1CstR is a persulfide-sensing member of the functionally diverse copper-sensitive operon repressor (CsoR) superfamily. While CstR regulates the bacterial response to hydrogen sulfide (H2S) and more oxidized reactive sulfur species (RSS) in Gram-positive pathogens, other dithiol-containing CsoR proteins respond to host derived Cu(I) toxicity, sometimes in the same bacterial cytoplasm, but without regulatory crosstalk in cells. It is not clear what prevents this crosstalk, nor the extent to which RSS sensors exhibit specificity over other oxidants. Here, we report a sequence similarity network (SSN) analysis of the entire CsoR superfamily, which together with the first crystallographic structure of a CstR and comprehensive mass spectrometry-based kinetic profiling experiments, reveal new insights into the molecular basis of RSS specificity in CstRs. We find that the more N-terminal cysteine is the attacking Cys in CstR and is far more nucleophilic than in a CsoR. Moreover, our CstR crystal structure is markedly asymmetric and chemical reactivity experiments reveal the functional impact of this asymmetry. Substitution of the Asn wedge between the resolving and the attacking thiol with Ala significantly decreases asymmetry in the crystal structure and markedly impacts the distribution of species, despite adopting the same global structure as the parent repressor. Companion NMR, SAXS and molecular dynamics simulations reveal that the structural and functional asymmetry can be traced to fast internal dynamics of the tetramer. Furthermore, this asymmetry is preserved in all CstRs and with all oxidants tested, giving rise to markedly distinct distributions of crosslinked products. Our exploration of the sequence, structural, and kinetic features that determine oxidant-specificity suggest that the product distribution upon RSS exposure is determined by internal flexibility.Fil: Fakhoury, Joseph N. Indiana University; Estados UnidosFil: Zhang, Yifan. Indiana University; Estados UnidosFil: Edmonds, Katherine A. Indiana University; Estados UnidosFil: Bringas, Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Luebke, Justin L. Indiana University; Estados UnidosFil: Gonzalez Gutierrez, Giovanni. Indiana University; Estados UnidosFil: Capdevila, Daiana Andrea. Indiana University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Giedroc, David Peter. Indiana University; Estados UnidosOxford University Press2021-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/167149Fakhoury, Joseph N; Zhang, Yifan; Edmonds, Katherine A; Bringas, Mauro; Luebke, Justin L; et al.; Functional asymmetry and chemical reactivity of CsoR family persulfide sensors; Oxford University Press; Nucleic Acids Research; 49; 21; 12-2021; 12556-125760305-10481362-4962CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkab1040info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/nar/article/49/21/12556/6424783info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:24:23Zoai:ri.conicet.gov.ar:11336/167149instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:24:24.034CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
title Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
spellingShingle Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
Fakhoury, Joseph N
TRANSCRIPTIONAL REGULATORS
RSS
STREPTOCOCCUS PNEUMONIAE
FUNCTIONAL ASSYMMMETRY
title_short Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
title_full Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
title_fullStr Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
title_full_unstemmed Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
title_sort Functional asymmetry and chemical reactivity of CsoR family persulfide sensors
dc.creator.none.fl_str_mv Fakhoury, Joseph N
Zhang, Yifan
Edmonds, Katherine A
Bringas, Mauro
Luebke, Justin L
Gonzalez Gutierrez, Giovanni
Capdevila, Daiana Andrea
Giedroc, David Peter
author Fakhoury, Joseph N
author_facet Fakhoury, Joseph N
Zhang, Yifan
Edmonds, Katherine A
Bringas, Mauro
Luebke, Justin L
Gonzalez Gutierrez, Giovanni
Capdevila, Daiana Andrea
Giedroc, David Peter
author_role author
author2 Zhang, Yifan
Edmonds, Katherine A
Bringas, Mauro
Luebke, Justin L
Gonzalez Gutierrez, Giovanni
Capdevila, Daiana Andrea
Giedroc, David Peter
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv TRANSCRIPTIONAL REGULATORS
RSS
STREPTOCOCCUS PNEUMONIAE
FUNCTIONAL ASSYMMMETRY
topic TRANSCRIPTIONAL REGULATORS
RSS
STREPTOCOCCUS PNEUMONIAE
FUNCTIONAL ASSYMMMETRY
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv CstR is a persulfide-sensing member of the functionally diverse copper-sensitive operon repressor (CsoR) superfamily. While CstR regulates the bacterial response to hydrogen sulfide (H2S) and more oxidized reactive sulfur species (RSS) in Gram-positive pathogens, other dithiol-containing CsoR proteins respond to host derived Cu(I) toxicity, sometimes in the same bacterial cytoplasm, but without regulatory crosstalk in cells. It is not clear what prevents this crosstalk, nor the extent to which RSS sensors exhibit specificity over other oxidants. Here, we report a sequence similarity network (SSN) analysis of the entire CsoR superfamily, which together with the first crystallographic structure of a CstR and comprehensive mass spectrometry-based kinetic profiling experiments, reveal new insights into the molecular basis of RSS specificity in CstRs. We find that the more N-terminal cysteine is the attacking Cys in CstR and is far more nucleophilic than in a CsoR. Moreover, our CstR crystal structure is markedly asymmetric and chemical reactivity experiments reveal the functional impact of this asymmetry. Substitution of the Asn wedge between the resolving and the attacking thiol with Ala significantly decreases asymmetry in the crystal structure and markedly impacts the distribution of species, despite adopting the same global structure as the parent repressor. Companion NMR, SAXS and molecular dynamics simulations reveal that the structural and functional asymmetry can be traced to fast internal dynamics of the tetramer. Furthermore, this asymmetry is preserved in all CstRs and with all oxidants tested, giving rise to markedly distinct distributions of crosslinked products. Our exploration of the sequence, structural, and kinetic features that determine oxidant-specificity suggest that the product distribution upon RSS exposure is determined by internal flexibility.
Fil: Fakhoury, Joseph N. Indiana University; Estados Unidos
Fil: Zhang, Yifan. Indiana University; Estados Unidos
Fil: Edmonds, Katherine A. Indiana University; Estados Unidos
Fil: Bringas, Mauro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Luebke, Justin L. Indiana University; Estados Unidos
Fil: Gonzalez Gutierrez, Giovanni. Indiana University; Estados Unidos
Fil: Capdevila, Daiana Andrea. Indiana University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Giedroc, David Peter. Indiana University; Estados Unidos
description CstR is a persulfide-sensing member of the functionally diverse copper-sensitive operon repressor (CsoR) superfamily. While CstR regulates the bacterial response to hydrogen sulfide (H2S) and more oxidized reactive sulfur species (RSS) in Gram-positive pathogens, other dithiol-containing CsoR proteins respond to host derived Cu(I) toxicity, sometimes in the same bacterial cytoplasm, but without regulatory crosstalk in cells. It is not clear what prevents this crosstalk, nor the extent to which RSS sensors exhibit specificity over other oxidants. Here, we report a sequence similarity network (SSN) analysis of the entire CsoR superfamily, which together with the first crystallographic structure of a CstR and comprehensive mass spectrometry-based kinetic profiling experiments, reveal new insights into the molecular basis of RSS specificity in CstRs. We find that the more N-terminal cysteine is the attacking Cys in CstR and is far more nucleophilic than in a CsoR. Moreover, our CstR crystal structure is markedly asymmetric and chemical reactivity experiments reveal the functional impact of this asymmetry. Substitution of the Asn wedge between the resolving and the attacking thiol with Ala significantly decreases asymmetry in the crystal structure and markedly impacts the distribution of species, despite adopting the same global structure as the parent repressor. Companion NMR, SAXS and molecular dynamics simulations reveal that the structural and functional asymmetry can be traced to fast internal dynamics of the tetramer. Furthermore, this asymmetry is preserved in all CstRs and with all oxidants tested, giving rise to markedly distinct distributions of crosslinked products. Our exploration of the sequence, structural, and kinetic features that determine oxidant-specificity suggest that the product distribution upon RSS exposure is determined by internal flexibility.
publishDate 2021
dc.date.none.fl_str_mv 2021-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/167149
Fakhoury, Joseph N; Zhang, Yifan; Edmonds, Katherine A; Bringas, Mauro; Luebke, Justin L; et al.; Functional asymmetry and chemical reactivity of CsoR family persulfide sensors; Oxford University Press; Nucleic Acids Research; 49; 21; 12-2021; 12556-12576
0305-1048
1362-4962
CONICET Digital
CONICET
url http://hdl.handle.net/11336/167149
identifier_str_mv Fakhoury, Joseph N; Zhang, Yifan; Edmonds, Katherine A; Bringas, Mauro; Luebke, Justin L; et al.; Functional asymmetry and chemical reactivity of CsoR family persulfide sensors; Oxford University Press; Nucleic Acids Research; 49; 21; 12-2021; 12556-12576
0305-1048
1362-4962
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkab1040
info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/nar/article/49/21/12556/6424783
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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