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Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans
Transcriptional adaptation to nitrate-dependent anabolism by Paracoccus denitrificans PD1222 was studied. A total of 74 genes were induced in cells grown with nitrate as N-source compared with ammonium, including nasTSABGHC and ntrBC genes. The nasT and nasS genes were cotranscribed, although nasT w...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424462/ https://www.ncbi.nlm.nih.gov/pubmed/28385879 http://dx.doi.org/10.1042/BCJ20170115 |
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author | Luque-Almagro, Victor M. Manso, Isabel Sullivan, Matthew J. Rowley, Gary Ferguson, Stuart J. Moreno-Vivián, Conrado Richardson, David J. Gates, Andrew J. Roldán, M. Dolores |
author_facet | Luque-Almagro, Victor M. Manso, Isabel Sullivan, Matthew J. Rowley, Gary Ferguson, Stuart J. Moreno-Vivián, Conrado Richardson, David J. Gates, Andrew J. Roldán, M. Dolores |
author_sort | Luque-Almagro, Victor M. |
collection | PubMed |
description | Transcriptional adaptation to nitrate-dependent anabolism by Paracoccus denitrificans PD1222 was studied. A total of 74 genes were induced in cells grown with nitrate as N-source compared with ammonium, including nasTSABGHC and ntrBC genes. The nasT and nasS genes were cotranscribed, although nasT was more strongly induced by nitrate than nasS. The nasABGHC genes constituted a transcriptional unit, which is preceded by a non-coding region containing hairpin structures involved in transcription termination. The nasTS and nasABGHC transcripts were detected at similar levels with nitrate or glutamate as N-source, but nasABGHC transcript was undetectable in ammonium-grown cells. The nitrite reductase NasG subunit was detected by two-dimensional polyacrylamide gel electrophoresis in cytoplasmic fractions from nitrate-grown cells, but it was not observed when either ammonium or glutamate was used as the N-source. The nasT mutant lacked both nasABGHC transcript and nicotinamide adenine dinucleotide (NADH)-dependent nitrate reductase activity. On the contrary, the nasS mutant showed similar levels of the nasABGHC transcript to the wild-type strain and displayed NasG protein and NADH–nitrate reductase activity with all N-sources tested, except with ammonium. Ammonium repression of nasABGHC was dependent on the Ntr system. The ntrBC and ntrYX genes were expressed at low levels regardless of the nitrogen source supporting growth. Mutational analysis of the ntrBCYX genes indicated that while ntrBC genes are required for nitrate assimilation, ntrYX genes can only partially restore growth on nitrate in the absence of ntrBC genes. The existence of a regulation mechanism for nitrate assimilation in P. denitrificans, by which nitrate induction operates at both transcriptional and translational levels, is proposed. |
format | Online Article Text |
id | pubmed-5424462 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-54244622017-05-22 Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans Luque-Almagro, Victor M. Manso, Isabel Sullivan, Matthew J. Rowley, Gary Ferguson, Stuart J. Moreno-Vivián, Conrado Richardson, David J. Gates, Andrew J. Roldán, M. Dolores Biochem J Research Articles Transcriptional adaptation to nitrate-dependent anabolism by Paracoccus denitrificans PD1222 was studied. A total of 74 genes were induced in cells grown with nitrate as N-source compared with ammonium, including nasTSABGHC and ntrBC genes. The nasT and nasS genes were cotranscribed, although nasT was more strongly induced by nitrate than nasS. The nasABGHC genes constituted a transcriptional unit, which is preceded by a non-coding region containing hairpin structures involved in transcription termination. The nasTS and nasABGHC transcripts were detected at similar levels with nitrate or glutamate as N-source, but nasABGHC transcript was undetectable in ammonium-grown cells. The nitrite reductase NasG subunit was detected by two-dimensional polyacrylamide gel electrophoresis in cytoplasmic fractions from nitrate-grown cells, but it was not observed when either ammonium or glutamate was used as the N-source. The nasT mutant lacked both nasABGHC transcript and nicotinamide adenine dinucleotide (NADH)-dependent nitrate reductase activity. On the contrary, the nasS mutant showed similar levels of the nasABGHC transcript to the wild-type strain and displayed NasG protein and NADH–nitrate reductase activity with all N-sources tested, except with ammonium. Ammonium repression of nasABGHC was dependent on the Ntr system. The ntrBC and ntrYX genes were expressed at low levels regardless of the nitrogen source supporting growth. Mutational analysis of the ntrBCYX genes indicated that while ntrBC genes are required for nitrate assimilation, ntrYX genes can only partially restore growth on nitrate in the absence of ntrBC genes. The existence of a regulation mechanism for nitrate assimilation in P. denitrificans, by which nitrate induction operates at both transcriptional and translational levels, is proposed. Portland Press Ltd. 2017-06-01 2017-05-10 /pmc/articles/PMC5424462/ /pubmed/28385879 http://dx.doi.org/10.1042/BCJ20170115 Text en © 2017 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Articles Luque-Almagro, Victor M. Manso, Isabel Sullivan, Matthew J. Rowley, Gary Ferguson, Stuart J. Moreno-Vivián, Conrado Richardson, David J. Gates, Andrew J. Roldán, M. Dolores Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title | Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title_full | Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title_fullStr | Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title_full_unstemmed | Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title_short | Transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier Paracoccus denitrificans |
title_sort | transcriptional and translational adaptation to aerobic nitrate anabolism in the denitrifier paracoccus denitrificans |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424462/ https://www.ncbi.nlm.nih.gov/pubmed/28385879 http://dx.doi.org/10.1042/BCJ20170115 |
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