Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i>
The gaseous hormone ethylene plays pivotal roles in plant growth and development. The rate-limiting enzyme of ethylene biosynthesis in seed plants is 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS). ACS proteins are encoded by a multigene family and the expression of <i>ACS</i>...
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MDPI AG
2024-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/5/2931 |
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| author | Xianglin Tang Ran Liu Yuanyuan Mei Dan Wang Kaixuan He Ning Ning Wang |
| author_facet | Xianglin Tang Ran Liu Yuanyuan Mei Dan Wang Kaixuan He Ning Ning Wang |
| author_sort | Xianglin Tang |
| collection | DOAJ |
| description | The gaseous hormone ethylene plays pivotal roles in plant growth and development. The rate-limiting enzyme of ethylene biosynthesis in seed plants is 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS). ACS proteins are encoded by a multigene family and the expression of <i>ACS</i> genes is highly regulated, especially at a post-translational level. AtACS7, the only type III ACS in <i>Arabidopsis</i>, is degraded in a 26S proteasome-dependent pathway. Here, by using liquid chromatography–mass spectrometry/mass spectrometry (LC-MS/MS) analysis, two lysine residues of AtACS7, lys285 (K285) and lys366 (K366), were revealed to be ubiquitin-modified in young, light-grown <i>Arabidopsis</i> seedlings but not in etiolated seedlings. Deubiquitylation-mimicking mutations of these residues significantly increased the stability of the AtACS7<sup>K285RK366R</sup> mutant protein in cell-free degradation assays. All results suggest that K285 and K366 are the major ubiquitination sites on AtACS7, providing deeper insights into the post-translational regulation of AtACS7 in <i>Arabidopsis</i>. |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-04-25T00:27:38Z |
| publishDate | 2024-03-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-56f9c32a015b4f2d942bd52ffb6af6512024-03-12T16:46:53ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-03-01255293110.3390/ijms25052931Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i>Xianglin Tang0Ran Liu1Yuanyuan Mei2Dan Wang3Kaixuan He4Ning Ning Wang5Tianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaTianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaTianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaTianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaTianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaTianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaThe gaseous hormone ethylene plays pivotal roles in plant growth and development. The rate-limiting enzyme of ethylene biosynthesis in seed plants is 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS). ACS proteins are encoded by a multigene family and the expression of <i>ACS</i> genes is highly regulated, especially at a post-translational level. AtACS7, the only type III ACS in <i>Arabidopsis</i>, is degraded in a 26S proteasome-dependent pathway. Here, by using liquid chromatography–mass spectrometry/mass spectrometry (LC-MS/MS) analysis, two lysine residues of AtACS7, lys285 (K285) and lys366 (K366), were revealed to be ubiquitin-modified in young, light-grown <i>Arabidopsis</i> seedlings but not in etiolated seedlings. Deubiquitylation-mimicking mutations of these residues significantly increased the stability of the AtACS7<sup>K285RK366R</sup> mutant protein in cell-free degradation assays. All results suggest that K285 and K366 are the major ubiquitination sites on AtACS7, providing deeper insights into the post-translational regulation of AtACS7 in <i>Arabidopsis</i>.https://www.mdpi.com/1422-0067/25/5/2931AtACS7ubiquitinationproteasomal degradationLC-MS/MS |
| spellingShingle | Xianglin Tang Ran Liu Yuanyuan Mei Dan Wang Kaixuan He Ning Ning Wang Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> International Journal of Molecular Sciences AtACS7 ubiquitination proteasomal degradation LC-MS/MS |
| title | Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> |
| title_full | Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> |
| title_fullStr | Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> |
| title_full_unstemmed | Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> |
| title_short | Identification of Key Ubiquitination Sites Involved in the Proteasomal Degradation of AtACS7 in <i>Arabidopsis</i> |
| title_sort | identification of key ubiquitination sites involved in the proteasomal degradation of atacs7 in i arabidopsis i |
| topic | AtACS7 ubiquitination proteasomal degradation LC-MS/MS |
| url | https://www.mdpi.com/1422-0067/25/5/2931 |
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