Phospho-Mutant Activity Assays Provide Evidence for the Negative Regulation of Transcriptional Regulator PRE1 by Phosphorylation.
Phospho-Mutant Activity Assays Provide Evidence for the Negative Regulation of Transcriptional Regulator PRE1 by Phosphorylation.
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DOI：
10.3390/ijms21239183
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发表时间：
2020-12-02
影响因子：
5.6
通讯作者：
Fan M
中科院分区：
文献类型：
--
作者：
Wang M;Tian Y;Han C;Zhou C;Bai MY;Fan M
paclobutrazol resistance （PRE）基因家族编码一组非典型螺旋-环-螺旋（HLH）蛋白，这些蛋白是整合广泛的环境和激素信号以调节植物生长和发育的主要枢纽。PRE1作为细胞延伸的正调节因子，通过隔离其抑制剂IBH1激活HBI1的DNA结合。此外，PRE1可以在Ser-46和Ser-67位点磷酸化，但这种磷酸化如何调节PRE1的功能尚不清楚。在这里，我们使用磷酸化突变活性测定来揭示Ser-67的磷酸化负调控PRE1对细胞伸长的功能。丝氨酸46的突变对PRE1的功能无显著影响，无论是磷酸化死丙氨酸还是磷酸化拟谷氨酸。然而，丝氨酸67突变为谷氨酸（PRE1S67E-Ox）而非丙氨酸（PRE1S67A-Ox）显著降低了PRE1对细胞延伸的促进作用。Ser-67向Glu-67的突变破坏了PRE1与IBH1的相互作用，导致PRE1无法抑制IBH1与HBI1的相互作用，失去了诱导后续细胞延长相关基因表达的能力。此外，我们发现PRE1-Ox和PRE1S67A-Ox对IBH1-Ox矮化表型均有抑制作用，但PRE1S67E-Ox对IBH1-Ox矮化表型无明显影响。我们的研究表明，Ser-67位点的磷酸化可负调控PRE1的活性。
The PACLOBUTRAZOL-RESISTANCE (PRE) gene family encodes a group of atypical helix-loop-helix (HLH) proteins that act as the major hub integrating a wide range of environmental and hormonal signals to regulate plant growth and development. PRE1, as a positive regulator of cell elongation, activates HBI1 DNA binding by sequestering its inhibitor IBH1. Furthermore, PRE1 can be phosphorylated at Ser-46 and Ser-67, but how this phosphorylation regulates the functions of PRE1 remains unclear. Here, we used a phospho-mutant activity assay to reveal that the phosphorylation at Ser-67 negatively regulates the functions of PRE1 on cell elongation. Both of mutations of serine 46, either to phospho-dead alanine or phospho-mimicking glutamic acid, had no significant effects on the functions of PRE1. However, the mutation of serine 67 to glutamic acid (PRE1S67E-Ox), but not alanine (PRE1S67A-Ox), significantly reduced the promoting effects of PRE1 on cell elongation. The mutation of Ser-67 to Glu-67 impaired the interaction of PRE1 with IBH1 and resulted in PRE1 failing to inhibit the interaction between IBH1 and HBI1, losing the ability to induce the expression of the subsequent cell elongation-related genes. Furthermore, we showed that PRE1-Ox and PRE1S67A-Ox both suppressed but PRE1S67E-Ox had no strong effects on the dwarf phenotypes of IBH1-Ox. Our study demonstrated that the PRE1 activity is negatively regulated by the phosphorylation at Ser-67.
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影响因子：
7
作者：
Ledent, V;Vervoort, M
通讯作者：
Vervoort, M
影响因子：
11.6
作者：
Leivar, Pablo;Tepperman, James M.;Quail, Peter H.
通讯作者：
Quail, Peter H.
DOI：
10.1073/pnas.1711723115
发表时间：
2017-12-26
影响因子：
11.1
作者：
Feng, Ying;Xu, Ping;Guo, Hongwei
通讯作者：
Guo, Hongwei
影响因子：
4.6
作者：
Bernhardt, C;Lee, MM;Schiefelbein, J
通讯作者：
Schiefelbein, J
DOI：
10.1073/pnas.96.10.5832
发表时间：
1999-05-11
影响因子：
11.1
作者：
Halliday, KJ;Hudson, M;Quail, PH
通讯作者：
Quail, PH