The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501
<i>Pseudomonas stutzeri</i> A1501, a plant-associated diazotrophic bacterium, prefers to conform to a nitrogen-fixing biofilm state under nitrogen-deficient conditions. The extracytoplasmic function (ECF) sigma factor AlgU is reported to play key roles in exopolysaccharide (EPS) producti...
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| Language: | English |
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MDPI AG
2022-05-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/13/5/867 |
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| author | Yahui Shao Changyan Yin Fanyang Lv Shanshan Jiang Shaoyu Wu Yueyue Han Wei Xue Yiyuan Ma Juan Zheng Yuhua Zhan Xiubin Ke Wei Lu Min Lin Liguo Shang Yongliang Yan |
| author_facet | Yahui Shao Changyan Yin Fanyang Lv Shanshan Jiang Shaoyu Wu Yueyue Han Wei Xue Yiyuan Ma Juan Zheng Yuhua Zhan Xiubin Ke Wei Lu Min Lin Liguo Shang Yongliang Yan |
| author_sort | Yahui Shao |
| collection | DOAJ |
| description | <i>Pseudomonas stutzeri</i> A1501, a plant-associated diazotrophic bacterium, prefers to conform to a nitrogen-fixing biofilm state under nitrogen-deficient conditions. The extracytoplasmic function (ECF) sigma factor AlgU is reported to play key roles in exopolysaccharide (EPS) production and biofilm formation in the <i>Pseudomonas</i> genus; however, the function of AlgU in <i>P. stutzeri</i> A1501 is still unclear. In this work, we mainly investigated the role of <i>algU</i> in EPS production, biofilm formation and nitrogenase activity in A1501. The <i>algU</i> mutant Δ<i>algU</i> showed a dramatic decrease both in the EPS production and the biofilm formation capabilities. In addition, the biofilm-based nitrogenase activity was reduced by 81.4% in the Δ<i>algU</i> mutant. The transcriptional level of <i>pslA</i>, a key Psl-like (a major EPS in A1501) synthesis-related gene, was almost completely inhibited in the <i>algU</i> mutant and was upregulated by 2.8-fold in the <i>algU</i>-overexpressing strain. A predicted AlgU-binding site was identified in the promoter region of <i>pslA</i>. The DNase I footprinting assays indicated that AlgU could directly bind to the <i>pslA</i> promoter, and β-galactosidase activity analysis further revealed mutations of the AlgU-binding boxes drastically reduced the transcriptional activity of the <i>pslA</i> promoter; moreover, we also demonstrated that AlgU was positively regulated by RpoN at the transcriptional level and negatively regulated by the RNA-binding protein RsmA at the posttranscriptional level. Taken together, these data suggest that AlgU promotes EPS production and nitrogen-fixing biofilm formation by directly activating the transcription of <i>pslA</i>, and the expression of AlgU is controlled by RpoN and RsmA at different regulatory levels. |
| first_indexed | 2024-03-10T03:49:44Z |
| format | Article |
| id | doaj.art-dec1691ee3b14b3f91cd1e627fb63111 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T03:49:44Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-dec1691ee3b14b3f91cd1e627fb631112023-11-23T11:10:45ZengMDPI AGGenes2073-44252022-05-0113586710.3390/genes13050867The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501Yahui Shao0Changyan Yin1Fanyang Lv2Shanshan Jiang3Shaoyu Wu4Yueyue Han5Wei Xue6Yiyuan Ma7Juan Zheng8Yuhua Zhan9Xiubin Ke10Wei Lu11Min Lin12Liguo Shang13Yongliang Yan14Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaSchool of Basic Medicine, GuangXi University of Chinese Medicine, Nanning 530200, ChinaBiotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China<i>Pseudomonas stutzeri</i> A1501, a plant-associated diazotrophic bacterium, prefers to conform to a nitrogen-fixing biofilm state under nitrogen-deficient conditions. The extracytoplasmic function (ECF) sigma factor AlgU is reported to play key roles in exopolysaccharide (EPS) production and biofilm formation in the <i>Pseudomonas</i> genus; however, the function of AlgU in <i>P. stutzeri</i> A1501 is still unclear. In this work, we mainly investigated the role of <i>algU</i> in EPS production, biofilm formation and nitrogenase activity in A1501. The <i>algU</i> mutant Δ<i>algU</i> showed a dramatic decrease both in the EPS production and the biofilm formation capabilities. In addition, the biofilm-based nitrogenase activity was reduced by 81.4% in the Δ<i>algU</i> mutant. The transcriptional level of <i>pslA</i>, a key Psl-like (a major EPS in A1501) synthesis-related gene, was almost completely inhibited in the <i>algU</i> mutant and was upregulated by 2.8-fold in the <i>algU</i>-overexpressing strain. A predicted AlgU-binding site was identified in the promoter region of <i>pslA</i>. The DNase I footprinting assays indicated that AlgU could directly bind to the <i>pslA</i> promoter, and β-galactosidase activity analysis further revealed mutations of the AlgU-binding boxes drastically reduced the transcriptional activity of the <i>pslA</i> promoter; moreover, we also demonstrated that AlgU was positively regulated by RpoN at the transcriptional level and negatively regulated by the RNA-binding protein RsmA at the posttranscriptional level. Taken together, these data suggest that AlgU promotes EPS production and nitrogen-fixing biofilm formation by directly activating the transcription of <i>pslA</i>, and the expression of AlgU is controlled by RpoN and RsmA at different regulatory levels.https://www.mdpi.com/2073-4425/13/5/867nitrogen fixationbiofilm formationthe <i>algU</i> genethe <i>pslA</i> gene<i>Pseudomonas stutzeri</i> |
| spellingShingle | Yahui Shao Changyan Yin Fanyang Lv Shanshan Jiang Shaoyu Wu Yueyue Han Wei Xue Yiyuan Ma Juan Zheng Yuhua Zhan Xiubin Ke Wei Lu Min Lin Liguo Shang Yongliang Yan The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 Genes nitrogen fixation biofilm formation the <i>algU</i> gene the <i>pslA</i> gene <i>Pseudomonas stutzeri</i> |
| title | The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 |
| title_full | The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 |
| title_fullStr | The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 |
| title_full_unstemmed | The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 |
| title_short | The Sigma Factor AlgU Regulates Exopolysaccharide Production and Nitrogen-Fixing Biofilm Formation by Directly Activating the Transcription of <i>pslA</i> in <i>Pseudomonas stutzeri</i> A1501 |
| title_sort | sigma factor algu regulates exopolysaccharide production and nitrogen fixing biofilm formation by directly activating the transcription of i psla i in i pseudomonas stutzeri i a1501 |
| topic | nitrogen fixation biofilm formation the <i>algU</i> gene the <i>pslA</i> gene <i>Pseudomonas stutzeri</i> |
| url | https://www.mdpi.com/2073-4425/13/5/867 |
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