Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice
Chrysanthemum (<i>Chrysanthemum morifolium</i> Ramat.) is one of the largest cut flowers in the world. Phosphate transporter Pht1 family member CmPht1;2 protein (CmPT2) plays an important role in response to low-phosphate (LP) stress in chrysanthemum. Post-translational modification (PTM...
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/3/2025 |
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| author | Jiayi Tang Chen Liu Yiqing Tan Jiafu Jiang Fadi Chen Guosheng Xiong Sumei Chen |
| author_facet | Jiayi Tang Chen Liu Yiqing Tan Jiafu Jiang Fadi Chen Guosheng Xiong Sumei Chen |
| author_sort | Jiayi Tang |
| collection | DOAJ |
| description | Chrysanthemum (<i>Chrysanthemum morifolium</i> Ramat.) is one of the largest cut flowers in the world. Phosphate transporter Pht1 family member CmPht1;2 protein (CmPT2) plays an important role in response to low-phosphate (LP) stress in chrysanthemum. Post-translational modification (PTM) can modulate the function of proteins in multiple ways. Here, we used yeast and rice systems to study the role of putative PTM in CmPT2 by determining the effect of mutation of key amino acid residues of putative glycosylation, phosphorylation, and myristoylation sites. We chose nine amino acid residues in the putative PTM sites and mutated them to alanine (A) (<i>Cmphts</i>). CmPT2 recovered the growth of yeast strain MB192 under LP conditions. However, G84A, G222A, T239A, Y242A, and N422A mutants could not grow normally under LP conditions. Analysis of phosphorus absorption kinetics showed that the Km of CmPT2 was 65.7 μM. Among the nine <i>Cmphts</i>, the expression of five with larger Km (124.4–397.5 μM) than CmPT2 was further evaluated in rice. Overexpression of <i>CmPT2</i>-OE increased plant height, effective panicle numbers, branch numbers, and yield compared with that of wild type ‘Wuyunjing No. 7’ (W7). Overexpression of <i>Cmphts</i>-OE led to decreased plant height and effective panicle numbers compared with that of the <i>CmPT2</i>-OE strain. The Pi content in roots of <i>CmPT2</i>-OE was higher than that of the W7 under both high (normal) phosphate (HP) and LP conditions. However, the Pi content in the leaves and roots was significantly lower in the N422A-OE strain than in the <i>CmPT2</i>-OE strain under both HP and LP conditions. Under LP conditions, the phosphorus starvation response (PSR) genes in <i>CmPT2</i>-OE were inhibited at the transcription level. The expression patterns of phosphorus-related genes in T239A, Y242A, and N422A-OE under LP conditions were different from those of <i>CmPT2</i>-OE. In conclusion, these five post-translational modification residues of CmPT2 play key roles in modulating the function of CmPT2. This work boosters our understanding of the function of phosphate transporters and provides genetic resources for improving the efficiency of phosphorus utilization in crop plants. |
| first_indexed | 2024-03-11T09:42:41Z |
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| spelling | doaj.art-3eaf7c08a74e407db1bddd105ba64b022023-11-16T16:51:14ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01243202510.3390/ijms24032025Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and RiceJiayi Tang0Chen Liu1Yiqing Tan2Jiafu Jiang3Fadi Chen4Guosheng Xiong5Sumei Chen6College of Horticulture, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Horticulture, Nanjing Agricultural University, Nanjing 210095, ChinaAcademy for Advanced Interdisciplinary Studies, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Horticulture, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Horticulture, Nanjing Agricultural University, Nanjing 210095, ChinaAcademy for Advanced Interdisciplinary Studies, Nanjing Agricultural University, Nanjing 210095, ChinaCollege of Horticulture, Nanjing Agricultural University, Nanjing 210095, ChinaChrysanthemum (<i>Chrysanthemum morifolium</i> Ramat.) is one of the largest cut flowers in the world. Phosphate transporter Pht1 family member CmPht1;2 protein (CmPT2) plays an important role in response to low-phosphate (LP) stress in chrysanthemum. Post-translational modification (PTM) can modulate the function of proteins in multiple ways. Here, we used yeast and rice systems to study the role of putative PTM in CmPT2 by determining the effect of mutation of key amino acid residues of putative glycosylation, phosphorylation, and myristoylation sites. We chose nine amino acid residues in the putative PTM sites and mutated them to alanine (A) (<i>Cmphts</i>). CmPT2 recovered the growth of yeast strain MB192 under LP conditions. However, G84A, G222A, T239A, Y242A, and N422A mutants could not grow normally under LP conditions. Analysis of phosphorus absorption kinetics showed that the Km of CmPT2 was 65.7 μM. Among the nine <i>Cmphts</i>, the expression of five with larger Km (124.4–397.5 μM) than CmPT2 was further evaluated in rice. Overexpression of <i>CmPT2</i>-OE increased plant height, effective panicle numbers, branch numbers, and yield compared with that of wild type ‘Wuyunjing No. 7’ (W7). Overexpression of <i>Cmphts</i>-OE led to decreased plant height and effective panicle numbers compared with that of the <i>CmPT2</i>-OE strain. The Pi content in roots of <i>CmPT2</i>-OE was higher than that of the W7 under both high (normal) phosphate (HP) and LP conditions. However, the Pi content in the leaves and roots was significantly lower in the N422A-OE strain than in the <i>CmPT2</i>-OE strain under both HP and LP conditions. Under LP conditions, the phosphorus starvation response (PSR) genes in <i>CmPT2</i>-OE were inhibited at the transcription level. The expression patterns of phosphorus-related genes in T239A, Y242A, and N422A-OE under LP conditions were different from those of <i>CmPT2</i>-OE. In conclusion, these five post-translational modification residues of CmPT2 play key roles in modulating the function of CmPT2. This work boosters our understanding of the function of phosphate transporters and provides genetic resources for improving the efficiency of phosphorus utilization in crop plants.https://www.mdpi.com/1422-0067/24/3/2025<i>CmPT2</i>yeast complementationpost-translational modificationrice |
| spellingShingle | Jiayi Tang Chen Liu Yiqing Tan Jiafu Jiang Fadi Chen Guosheng Xiong Sumei Chen Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice International Journal of Molecular Sciences <i>CmPT2</i> yeast complementation post-translational modification rice |
| title | Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice |
| title_full | Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice |
| title_fullStr | Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice |
| title_full_unstemmed | Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice |
| title_short | Five Post-Translational Modification Residues of CmPT2 Play Key Roles in Yeast and Rice |
| title_sort | five post translational modification residues of cmpt2 play key roles in yeast and rice |
| topic | <i>CmPT2</i> yeast complementation post-translational modification rice |
| url | https://www.mdpi.com/1422-0067/24/3/2025 |
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