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Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense

Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has...

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Autores principales: Sotomaior, P., Araújo, L.M., Nishikawa, C.Y., Huergo, L.F., Monteiro, R.A., Pedrosa, F.O., Chubatsu, L.S., Souza, E.M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Medicina Tropical 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3854208/
https://www.ncbi.nlm.nih.gov/pubmed/22983183
http://dx.doi.org/10.1590/S0100-879X2012007500146
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author Sotomaior, P.
Araújo, L.M.
Nishikawa, C.Y.
Huergo, L.F.
Monteiro, R.A.
Pedrosa, F.O.
Chubatsu, L.S.
Souza, E.M.
author_facet Sotomaior, P.
Araújo, L.M.
Nishikawa, C.Y.
Huergo, L.F.
Monteiro, R.A.
Pedrosa, F.O.
Chubatsu, L.S.
Souza, E.M.
author_sort Sotomaior, P.
collection PubMed
description Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.
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spelling pubmed-38542082013-12-16 Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense Sotomaior, P. Araújo, L.M. Nishikawa, C.Y. Huergo, L.F. Monteiro, R.A. Pedrosa, F.O. Chubatsu, L.S. Souza, E.M. Braz J Med Biol Res Short Communication Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate. Sociedade Brasileira de Medicina Tropical 2012-09-21 /pmc/articles/PMC3854208/ /pubmed/22983183 http://dx.doi.org/10.1590/S0100-879X2012007500146 Text en http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Short Communication
Sotomaior, P.
Araújo, L.M.
Nishikawa, C.Y.
Huergo, L.F.
Monteiro, R.A.
Pedrosa, F.O.
Chubatsu, L.S.
Souza, E.M.
Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title_full Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title_fullStr Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title_full_unstemmed Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title_short Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense
title_sort effect of atp and 2-oxoglutarate on the in vitro interaction between the nifa gaf domain and the glnb protein of azospirillum brasilense
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3854208/
https://www.ncbi.nlm.nih.gov/pubmed/22983183
http://dx.doi.org/10.1590/S0100-879X2012007500146
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