Secreted citrate serves as iron carrier for the marine pathogen photobacterium damselae subsp damselae
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Título: | Secreted citrate serves as iron carrier for the marine pathogen photobacterium damselae subsp damselae |
Autor/a: | Balado Dacosta, Miguel Puentes Corral, Beatriz Couceiro López, Lucía Fuentes Monteverde, Juan Carlos Rodríguez, Jaime Rodríguez Osorio, Carlos Jiménez, Carlos Lemos Ramos, Manuel Luis |
Centro/Departamento: | Universidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía Universidade de Santiago de Compostela. Instituto de Acuicultura |
Palabras chave: | Photobacterium damselae | Citrate | Iron uptake | Siderophores | Vibrioferrin | |
Data: | 2017 |
Editor: | Frontiers Media |
Cita bibliográfica: | Balado M, Puentes B, Couceiro L, Fuentes-Monteverde JC, Rodríguez J, Osorio CR, Jiménez C and Lemos ML (2017) Secreted Citrate Serves as Iron Carrier for the Marine Pathogen Photobacterium damselae subsp damselae. Front. Cell. Infect. Microbiol. 7:361. doi: 10.3389/fcimb.2017.00361 |
Resumo: | Photobacterium damselae subsp damselae (Pdd) is a Vibrionaceae that has a wide pathogenic potential against many marine animals and also against humans. Some strains of this bacterium acquire iron through the siderophore vibrioferrin. However, there are virulent strains that do not produce vibrioferrin, but they still give a strong positive reaction in the CAS test for siderophore production. In an in silico search on the genome sequences of this type of strains we could not find any ORF which could be related to a siderophore system. To identify genes that could encode a siderophore-mediated iron acquisition system we used a mini-Tn10 transposon random mutagenesis approach. From more than 1,400 mutants examined, we could isolate a mutant (BP53) that showed a strong CAS reaction independently of the iron levels of the medium. In this mutant the transposon was inserted into the idh gene, which encodes an isocitrate dehydrogenase that participates in the tricarboxylic acid cycle. The mutant did not show any growth impairment in rich or minimal media, but it accumulated a noticeable amount of citrate (around 7 mM) in the culture medium, irrespective of the iron levels. The parental strain accumulated citrate, but in an iron-regulated fashion, being citrate levels 5–6 times higher under iron restricted conditions. In addition, a null mutant deficient in citrate synthase showed an impairment for growth at high concentrations of iron chelators, and showed almost no reaction in the CAS test. Chemical analysis by liquid chromatography of the iron-restricted culture supernatants resulted in a CAS-positive fraction with biological activity as siderophore. HPLC purification of that fraction yielded a pure compound which was identified as citrate from its MS and NMR spectral data. Although the production of another citrate-based compound with siderophore activity cannot be ruled out, our results suggest that Pdd secretes endogenous citrate and use it for iron scavenging from the cell environment |
Versión do editor: | https://doi.org/10.3389/fcimb.2017.00361 |
URI: | http://hdl.handle.net/10347/22783 |
DOI: | 10.3389/fcimb.2017.00361 |
E-ISSN: | 2235-2988 |
Dereitos: | Copyright © 2017 Balado, Puentes, Couceiro, Fuentes-Monteverde, Rodríguez, Osorio, Jiménez and Lemos. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms |
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Copyright © 2017 Balado, Puentes, Couceiro, Fuentes-Monteverde, Rodríguez, Osorio, Jiménez and Lemos. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms
Copyright © 2017 Balado, Puentes, Couceiro, Fuentes-Monteverde, Rodríguez, Osorio, Jiménez and Lemos. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms