Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells
Plant defense and growth rely on multiple transcriptional factors (TFs). Repression of shoot growth (RSG) is a TF belonging to a bZIP family in tobacco, known to be involved in plant gibberellin feedback regulation by inducing the expression of key genes. The tobacco calcium-dependent protein kinase...
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MDPI AG
2022-08-01
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| Online Access: | https://www.mdpi.com/1422-0067/23/16/8941 |
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| author | Luyao Wang Ying Gui Bingye Yang Wenpan Dong Peiling Xu Fangjie Si Wei Yang Yuming Luo Jianhua Guo Dongdong Niu Chunhao Jiang |
| author_facet | Luyao Wang Ying Gui Bingye Yang Wenpan Dong Peiling Xu Fangjie Si Wei Yang Yuming Luo Jianhua Guo Dongdong Niu Chunhao Jiang |
| author_sort | Luyao Wang |
| collection | DOAJ |
| description | Plant defense and growth rely on multiple transcriptional factors (TFs). Repression of shoot growth (RSG) is a TF belonging to a bZIP family in tobacco, known to be involved in plant gibberellin feedback regulation by inducing the expression of key genes. The tobacco calcium-dependent protein kinase CDPK1 was reported to interact with RSG and manipulate its intracellular localization by phosphorylating Ser-114 of RSG previously. Here, we identified tobacco mitogen-activated protein kinase 3 (NtMPK3) as an RSG-interacting protein kinase. Moreover, the mutation of the predicted MAPK-associated phosphorylation site of RSG (Thr-30, Ser-74, and Thr-135) significantly altered the intracellular localization of the NtMPK3-RSG interaction complex. Nuclear transport of RSG and its amino acid mutants (T30A and S74A) were observed after being treated with plant defense elicitor peptide flg22 within 5 min, and the two mutated RSG swiftly re-localized in tobacco cytoplasm within 30 min. In addition, triple-point mutation of RSG (T30A/S74A/T135A) mimics constant unphosphorylated status, and is predominantly localized in tobacco cytoplasm. RSG (T30A/S74A/T135A) showed no re-localization effect under the treatments of flg22, <i>B. cereus</i> AR156, or GA<sub>3</sub>, and over-expression of this mutant in tobacco resulted in lower expression levels of downstream gene <i>GA20ox1</i>. Our results suggest that MAPK-associated phosphorylation sites of RSG regulate its localization in tobacco, and that constant unphosphorylation of RSG in Thr-30, Ser-74, and Thr-135 keeps RSG predominantly localized in cytoplasm. |
| first_indexed | 2024-03-09T04:20:55Z |
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| language | English |
| last_indexed | 2024-03-09T04:20:55Z |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-65ae605d32894ef48a99e2fb6f4d59e82023-12-03T13:47:24ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012316894110.3390/ijms23168941Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant CellsLuyao Wang0Ying Gui1Bingye Yang2Wenpan Dong3Peiling Xu4Fangjie Si5Wei Yang6Yuming Luo7Jianhua Guo8Dongdong Niu9Chunhao Jiang10Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaJiangsu Key Laboratory for Eco-Agricultural Biotechnology around Hongze Lake, Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Protection, Huaiyin Normal University, Huai’an 223300, ChinaJiangsu Key Laboratory for Eco-Agricultural Biotechnology around Hongze Lake, Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Protection, Huaiyin Normal University, Huai’an 223300, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaDepartment of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, ChinaPlant defense and growth rely on multiple transcriptional factors (TFs). Repression of shoot growth (RSG) is a TF belonging to a bZIP family in tobacco, known to be involved in plant gibberellin feedback regulation by inducing the expression of key genes. The tobacco calcium-dependent protein kinase CDPK1 was reported to interact with RSG and manipulate its intracellular localization by phosphorylating Ser-114 of RSG previously. Here, we identified tobacco mitogen-activated protein kinase 3 (NtMPK3) as an RSG-interacting protein kinase. Moreover, the mutation of the predicted MAPK-associated phosphorylation site of RSG (Thr-30, Ser-74, and Thr-135) significantly altered the intracellular localization of the NtMPK3-RSG interaction complex. Nuclear transport of RSG and its amino acid mutants (T30A and S74A) were observed after being treated with plant defense elicitor peptide flg22 within 5 min, and the two mutated RSG swiftly re-localized in tobacco cytoplasm within 30 min. In addition, triple-point mutation of RSG (T30A/S74A/T135A) mimics constant unphosphorylated status, and is predominantly localized in tobacco cytoplasm. RSG (T30A/S74A/T135A) showed no re-localization effect under the treatments of flg22, <i>B. cereus</i> AR156, or GA<sub>3</sub>, and over-expression of this mutant in tobacco resulted in lower expression levels of downstream gene <i>GA20ox1</i>. Our results suggest that MAPK-associated phosphorylation sites of RSG regulate its localization in tobacco, and that constant unphosphorylation of RSG in Thr-30, Ser-74, and Thr-135 keeps RSG predominantly localized in cytoplasm.https://www.mdpi.com/1422-0067/23/16/8941MAPKphosphorylationRSGabiotic stressprotein interaction |
| spellingShingle | Luyao Wang Ying Gui Bingye Yang Wenpan Dong Peiling Xu Fangjie Si Wei Yang Yuming Luo Jianhua Guo Dongdong Niu Chunhao Jiang Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells International Journal of Molecular Sciences MAPK phosphorylation RSG abiotic stress protein interaction |
| title | Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells |
| title_full | Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells |
| title_fullStr | Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells |
| title_full_unstemmed | Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells |
| title_short | Mitogen-Activated Protein Kinases Associated Sites of Tobacco Repression of Shoot Growth Regulates Its Localization in Plant Cells |
| title_sort | mitogen activated protein kinases associated sites of tobacco repression of shoot growth regulates its localization in plant cells |
| topic | MAPK phosphorylation RSG abiotic stress protein interaction |
| url | https://www.mdpi.com/1422-0067/23/16/8941 |
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