RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins

The plant pathogenic bacterium Ralstonia solanacearum injects more than 70 effector proteins (virulence factors) into the host plant cells via the needle-like structure of a type III secretion system. The type III secretion system effector proteins manipulate host regulatory networks to suppress def...

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Autores: Fujiwara, Shoko, Kawazoe, Tomoki, Ohnishi, Kouhei, Kitagawa, Takao, Popa, Crina Mihaela, Valls i Matheu, Marc, Genin, Stéphane, Nakamura, Kazuyuki, Kuramitsu, Yasuhiro, Tanaka, Naotaka, Tabuchi, Mitsuaki
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/103386
Acceso en línea:https://hdl.handle.net/2445/103386
Access Level:acceso abierto
Palabra clave:Saccharomyces cerevisiae
Botànica
Proteïnes
Reacció d'oxidació-reducció
Botany
Proteins
Oxidation-reduction reaction
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spelling RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxinsFujiwara, ShokoKawazoe, TomokiOhnishi, KouheiKitagawa, TakaoPopa, Crina MihaelaValls i Matheu, MarcGenin, StéphaneNakamura, KazuyukiKuramitsu, YasuhiroTanaka, NaotakaTabuchi, MitsuakiSaccharomyces cerevisiaeBotànicaProteïnesReacció d'oxidació-reduccióSaccharomyces cerevisiaeBotanyProteinsOxidation-reduction reactionThe plant pathogenic bacterium Ralstonia solanacearum injects more than 70 effector proteins (virulence factors) into the host plant cells via the needle-like structure of a type III secretion system. The type III secretion system effector proteins manipulate host regulatory networks to suppress defense responses with diverse molecular activities. Uncovering the molecular function of these effectors is essential for a mechanistic understanding of R. solanacearum pathogenicity. However, few of the effectors from R. solanacearum have been functionally characterized, and their plant targets remain largely unknown. Here, we show that the ChaC domain-containing effector RipAY/RSp1022 from R. solanacearum exhibits γ-glutamyl cyclotransferase (GGCT) activity to degrade the major intracellular redox buffer, glutathione. Heterologous expression of RipAY, but not other ChaC family proteins conserved in various organisms, caused growth inhibition of yeast Saccharomyces cerevisiae, and the intracellular glutathione level was decreased to ∼30% of the normal level following expression of RipAY in yeast. Although active site mutants of GGCT activity were non-toxic, the addition of glutathione did not reverse the toxicity, suggesting that the toxicity might be a consequence of activity against other γ-glutamyl compounds. Intriguingly, RipAY protein purified from a bacterial expression system did not exhibit any GGCT activity, whereas it exhibited robust GGCT activity upon its interaction with eukaryotic thioredoxins, which are important for intracellular redox homeostasis during bacterial infection in plants. Our results suggest that RipAY has evolved to sense the host intracellular redox environment, which triggers its enzymatic activity to create a favorable environment for R. solanacearum infection.American Society for Biochemistry and Molecular Biology2016info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/103386Articles publicats en revistes (Genètica, Microbiologia i Estadística)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1074/jbc.M115.678953Journal of Biological Chemistry, 2016, vol. 291, num. 13, p. 6813-6830https://doi.org/10.1074/jbc.M115.678953(c) American Society for Biochemistry and Molecular Biology, 2016info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1033862026-05-27T06:46:51Z
dc.title.none.fl_str_mv RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
title RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
spellingShingle RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
Fujiwara, Shoko
Saccharomyces cerevisiae
Botànica
Proteïnes
Reacció d'oxidació-reducció
Saccharomyces cerevisiae
Botany
Proteins
Oxidation-reduction reaction
title_short RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
title_full RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
title_fullStr RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
title_full_unstemmed RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
title_sort RipAY, a plant pathogen effector protein exhibits robust γ-glutamyl cyclotransferase activity when stimulated by eukaryotic thioredoxins
dc.creator.none.fl_str_mv Fujiwara, Shoko
Kawazoe, Tomoki
Ohnishi, Kouhei
Kitagawa, Takao
Popa, Crina Mihaela
Valls i Matheu, Marc
Genin, Stéphane
Nakamura, Kazuyuki
Kuramitsu, Yasuhiro
Tanaka, Naotaka
Tabuchi, Mitsuaki
author Fujiwara, Shoko
author_facet Fujiwara, Shoko
Kawazoe, Tomoki
Ohnishi, Kouhei
Kitagawa, Takao
Popa, Crina Mihaela
Valls i Matheu, Marc
Genin, Stéphane
Nakamura, Kazuyuki
Kuramitsu, Yasuhiro
Tanaka, Naotaka
Tabuchi, Mitsuaki
author_role author
author2 Kawazoe, Tomoki
Ohnishi, Kouhei
Kitagawa, Takao
Popa, Crina Mihaela
Valls i Matheu, Marc
Genin, Stéphane
Nakamura, Kazuyuki
Kuramitsu, Yasuhiro
Tanaka, Naotaka
Tabuchi, Mitsuaki
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Saccharomyces cerevisiae
Botànica
Proteïnes
Reacció d'oxidació-reducció
Saccharomyces cerevisiae
Botany
Proteins
Oxidation-reduction reaction
topic Saccharomyces cerevisiae
Botànica
Proteïnes
Reacció d'oxidació-reducció
Saccharomyces cerevisiae
Botany
Proteins
Oxidation-reduction reaction
description The plant pathogenic bacterium Ralstonia solanacearum injects more than 70 effector proteins (virulence factors) into the host plant cells via the needle-like structure of a type III secretion system. The type III secretion system effector proteins manipulate host regulatory networks to suppress defense responses with diverse molecular activities. Uncovering the molecular function of these effectors is essential for a mechanistic understanding of R. solanacearum pathogenicity. However, few of the effectors from R. solanacearum have been functionally characterized, and their plant targets remain largely unknown. Here, we show that the ChaC domain-containing effector RipAY/RSp1022 from R. solanacearum exhibits γ-glutamyl cyclotransferase (GGCT) activity to degrade the major intracellular redox buffer, glutathione. Heterologous expression of RipAY, but not other ChaC family proteins conserved in various organisms, caused growth inhibition of yeast Saccharomyces cerevisiae, and the intracellular glutathione level was decreased to ∼30% of the normal level following expression of RipAY in yeast. Although active site mutants of GGCT activity were non-toxic, the addition of glutathione did not reverse the toxicity, suggesting that the toxicity might be a consequence of activity against other γ-glutamyl compounds. Intriguingly, RipAY protein purified from a bacterial expression system did not exhibit any GGCT activity, whereas it exhibited robust GGCT activity upon its interaction with eukaryotic thioredoxins, which are important for intracellular redox homeostasis during bacterial infection in plants. Our results suggest that RipAY has evolved to sense the host intracellular redox environment, which triggers its enzymatic activity to create a favorable environment for R. solanacearum infection.
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://hdl.handle.net/2445/103386
url https://hdl.handle.net/2445/103386
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1074/jbc.M115.678953
Journal of Biological Chemistry, 2016, vol. 291, num. 13, p. 6813-6830
https://doi.org/10.1074/jbc.M115.678953
dc.rights.none.fl_str_mv (c) American Society for Biochemistry and Molecular Biology, 2016
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Society for Biochemistry and Molecular Biology, 2016
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv Articles publicats en revistes (Genètica, Microbiologia i Estadística)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301603

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