Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex

Cyanobacteria, phototrophic organisms that perform oxygenic photosynthesis, perceive nitrogen status by sensing 2-oxoglutarate levels. PII, a widespread signaling protein, senses and transduces nitrogen and energy status to target proteins, regulating metabolism and gene expression. In cyanobacteria...

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Detalles Bibliográficos
Autores: Labella, Jose I., Obrebska, Anna, Espinosa, Javier, Salinas, Paloma, Forcada-Nadal, Alicia, Tremiño, Lorena, Rubio, Vicente, Contreras, Asunción
Tipo de recurso: artículo
Fecha de publicación:2016
País:España
Institución:Universitat Politècnica de València (UPV)
Repositorio:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglés
OAI Identifier:oai:riunet.upv.es:10251/203997
Acceso en línea:https://riunet.upv.es/handle/10251/203997
Access Level:acceso abierto
Palabra clave:PlmA
PII
PipX
Cyanobacteria
Nitrogen regulation
Signaling
GntR
Three hybrid interactions
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network_name_str España
repository_id_str
dc.title.none.fl_str_mv Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
title Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
spellingShingle Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
Labella, Jose I.
PlmA
PII
PipX
Cyanobacteria
Nitrogen regulation
Signaling
GntR
Three hybrid interactions
title_short Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
title_full Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
title_fullStr Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
title_full_unstemmed Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
title_sort Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX Complex
dc.creator.none.fl_str_mv Labella, Jose I.
Obrebska, Anna
Espinosa, Javier
Salinas, Paloma
Forcada-Nadal, Alicia
Tremiño, Lorena
Rubio, Vicente
Contreras, Asunción
author Labella, Jose I.
author_facet Labella, Jose I.
Obrebska, Anna
Espinosa, Javier
Salinas, Paloma
Forcada-Nadal, Alicia
Tremiño, Lorena
Rubio, Vicente
Contreras, Asunción
author_role author
author2 Obrebska, Anna
Espinosa, Javier
Salinas, Paloma
Forcada-Nadal, Alicia
Tremiño, Lorena
Rubio, Vicente
Contreras, Asunción
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Generalitat Valenciana
Ministerio de Economía y Competitividad
Repositorio Institucional de la Universitat Politècnica de València Riunet
dc.subject.none.fl_str_mv PlmA
PII
PipX
Cyanobacteria
Nitrogen regulation
Signaling
GntR
Three hybrid interactions
topic PlmA
PII
PipX
Cyanobacteria
Nitrogen regulation
Signaling
GntR
Three hybrid interactions
description Cyanobacteria, phototrophic organisms that perform oxygenic photosynthesis, perceive nitrogen status by sensing 2-oxoglutarate levels. PII, a widespread signaling protein, senses and transduces nitrogen and energy status to target proteins, regulating metabolism and gene expression. In cyanobacteria, under conditions of low 2-oxoglutarate, PII forms complexes with the enzyme N-acetyl glutamate kinase, increasing arginine biosynthesis, and with PII-interacting protein X (PipX), making PipX unavailable for binding and co-activation of the nitrogen regulator NtcA. Both the PII-PipX complex structure and in vivo functional data suggested that this complex, as such, could have regulatory functions in addition to PipX sequestration. To investigate this possibilitywe performed yeast three-hybrid screening of genomic libraries from Synechococcus elongatus PCC7942, searching for proteins interacting simultaneously with PII and PipX. The only prey clone found in the search expressed PlmA, a member of the GntR family of transcriptional regulators proven here by gel filtration to be homodimeric. Interactions analyses further confirmed the simultaneous requirement of PII and PipX, and showed that the PlmA contacts involve PipX elements exposed in the PII-PipX complex, specifically the C-terminal helices and one residue of the tudor-like body. In contrast, PII appears not to interact directly with PlmA, possibly being needed indirectly, to induce an extended conformation of the C-terminal helices of PipX and for modulating the surface polarity at the PII-PipX boundary, two elements that appear crucial for PlmA binding. Attempts to inactive plmA confirmed that this gene is essential in S. elongatus. Western blot assays revealed that S. elongatus PlmA, irrespective of the nitrogen regime, is a relatively abundant transcriptional regulator, suggesting the existence of a large PlmA regulon. In silico studies showed that PlmA is universally and exclusively found in cyanobacteria. Based on interaction data, on the relative amounts of the proteins involved in PII-PipX-PlmA complexes, determined in western assays, and on the restrictions imposed by the symmetries of trimeric PII and dimeric PlmA molecules, a structural and regulatory model for PlmA function is discussed in the context of the cyanobacterial nitrogen interaction network.
publishDate 2016
dc.date.none.fl_str_mv 2016
2016-10-28
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://riunet.upv.es/handle/10251/203997
url https://riunet.upv.es/handle/10251/203997
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 BFU2014-58229-P UNA MIRADA MOLECULAR AL CONTROL DE LA DETOXIFICACION DE AMONIO Y A SUS PATOLOGIAS Y ERRORES CONGENITOS, A LA SEÑALIZACION POR NITROGENO. EN BUSCA DEL PAPEL DE LA PROTEINA CUTA
Ministerio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 BFU2015-66360-P MULTIFUNCIONALIDAD DE PIPX Y REDES DE SEÑALIZACION EN LA CIANOBACTERIA MODELO SYNECHOCOCCUS ELONGATUS SP. PCC 7942
Generalitat Valenciana https://doi.org/10.13039/501100003359 GV%2F2014%2F073
Generalitat Valenciana https://doi.org/10.13039/501100003359 PROMETEOII%2F2014%2F029
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media SA
publisher.none.fl_str_mv Frontiers Media SA
dc.source.none.fl_str_mv reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
instname:Universitat Politècnica de València (UPV)
instname_str Universitat Politècnica de València (UPV)
reponame_str RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
collection RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Expanding the Cyanobacterial Nitrogen Regulatory Network: The GntR-Like Regulator PlmA Interacts with the PII-PipX ComplexLabella, Jose I.Obrebska, AnnaEspinosa, JavierSalinas, PalomaForcada-Nadal, AliciaTremiño, LorenaRubio, VicenteContreras, AsunciónPlmAPIIPipXCyanobacteriaNitrogen regulationSignalingGntRThree hybrid interactionsCyanobacteria, phototrophic organisms that perform oxygenic photosynthesis, perceive nitrogen status by sensing 2-oxoglutarate levels. PII, a widespread signaling protein, senses and transduces nitrogen and energy status to target proteins, regulating metabolism and gene expression. In cyanobacteria, under conditions of low 2-oxoglutarate, PII forms complexes with the enzyme N-acetyl glutamate kinase, increasing arginine biosynthesis, and with PII-interacting protein X (PipX), making PipX unavailable for binding and co-activation of the nitrogen regulator NtcA. Both the PII-PipX complex structure and in vivo functional data suggested that this complex, as such, could have regulatory functions in addition to PipX sequestration. To investigate this possibilitywe performed yeast three-hybrid screening of genomic libraries from Synechococcus elongatus PCC7942, searching for proteins interacting simultaneously with PII and PipX. The only prey clone found in the search expressed PlmA, a member of the GntR family of transcriptional regulators proven here by gel filtration to be homodimeric. Interactions analyses further confirmed the simultaneous requirement of PII and PipX, and showed that the PlmA contacts involve PipX elements exposed in the PII-PipX complex, specifically the C-terminal helices and one residue of the tudor-like body. In contrast, PII appears not to interact directly with PlmA, possibly being needed indirectly, to induce an extended conformation of the C-terminal helices of PipX and for modulating the surface polarity at the PII-PipX boundary, two elements that appear crucial for PlmA binding. Attempts to inactive plmA confirmed that this gene is essential in S. elongatus. Western blot assays revealed that S. elongatus PlmA, irrespective of the nitrogen regime, is a relatively abundant transcriptional regulator, suggesting the existence of a large PlmA regulon. In silico studies showed that PlmA is universally and exclusively found in cyanobacteria. Based on interaction data, on the relative amounts of the proteins involved in PII-PipX-PlmA complexes, determined in western assays, and on the restrictions imposed by the symmetries of trimeric PII and dimeric PlmA molecules, a structural and regulatory model for PlmA function is discussed in the context of the cyanobacterial nitrogen interaction network.This work was supported by grants BFU2015-66360-P to AC and BFU2014-58229-P to VR from the Spanish Ministry of Economy and Competitivity. AO was the recipient of Grisolia Fellowship from Consellería d Educació of the Valencian Government and AF-N and LT held FPI fellowships/contracts from Ministry of Economy and Competitivity. JE and VR were supported by grants GV/2014/073 and PrometeoII/2014/029, respectively, from the Consellería d Educació of the Valencian Government.Frontiers Media SAGeneralitat ValencianaMinisterio de Economía y CompetitividadRepositorio Institucional de la Universitat Politècnica de València Riunet20162016-10-28journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://riunet.upv.es/handle/10251/203997reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengMinisterio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 BFU2014-58229-P UNA MIRADA MOLECULAR AL CONTROL DE LA DETOXIFICACION DE AMONIO Y A SUS PATOLOGIAS Y ERRORES CONGENITOS, A LA SEÑALIZACION POR NITROGENO. EN BUSCA DEL PAPEL DE LA PROTEINA CUTAMinisterio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 BFU2015-66360-P MULTIFUNCIONALIDAD DE PIPX Y REDES DE SEÑALIZACION EN LA CIANOBACTERIA MODELO SYNECHOCOCCUS ELONGATUS SP. PCC 7942Generalitat Valenciana https://doi.org/10.13039/501100003359 GV%2F2014%2F073Generalitat Valenciana https://doi.org/10.13039/501100003359 PROMETEOII%2F2014%2F029open accesshttp://purl.org/coar/access_right/c_abf2Reconocimiento (by)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/2039972026-06-13T07:49:27Z
score 15,300719

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