Structural and functional insights into the mechanism of action of plant borate transporters.
Structural and functional insights into the mechanism of action of plant borate transporters.
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DOI：
10.1038/s41598-021-91763-6
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发表时间：
2021-06-10
影响因子：
4.6
通讯作者：
Byrne B
中科院分区：
文献类型：
--
作者：
Saouros S;Mohan TC;Cecchetti C;Lehmann S;Barrit JD;Scull NJ;Simpson P;Alguel Y;Cameron AD;Jones AME;Byrne B
硼在植物生长和发育中具有重要作用。 BOR 蛋白是硼的主动吸收和分配以及细胞内硼浓度调节的关键。然而，它们的作用机制仍然缺乏研究。 BOR 蛋白是人类 SLC4 转运蛋白家族的同源物，其中包括经过深入研究的哺乳动物转运蛋白，例如人类阴离子交换蛋白 1 (hAE1)。在这里，我们生成了拟南芥 BOR1 (AtBOR1) 变体，其基础是 (i) 已知的 hAE1 (S466R、A500R) 致病突变和 (ii) 在酵母 BOR 蛋白 ScBOR1p 中发现的功能丧失突变 (D311A)。 AtBOR1 变体在酵母中表达并定位于质膜，但 S466R 和 A500R 的表达量均低于 WT AtBOR1 和 D311A。 D311A、S466R 和 A500R 突变导致酵母 bor1p 敲除菌株中硼酸盐流出活性丧失。含有这三个单独突变的拟南芥植物在低硼条件下在土壤中表现出显着降低的生长表型。这些数据证实了 D311 在蛋白质功能中的重要作用，并表明与 hAE1 中致病突变相当的突变对 AtBOR1 具有重大影响。我们还获得了来自稻的 BOR 蛋白 OsBOR3 的低分辨率冷冻电镜结构，该蛋白缺乏 30 个 C 端氨基酸残基。这种结构证实了之前观察到的相关蛋白质的门和核心结构域组织，并且强烈暗示了向内的构象。
Boron has essential roles in plant growth and development. BOR proteins are key in the active uptake and distribution of boron, and regulation of intracellular boron concentrations. However, their mechanism of action remains poorly studied. BOR proteins are homologues of the human SLC4 family of transporters, which includes well studied mammalian transporters such as the human Anion Exchanger 1 (hAE1). Here we generated Arabidopsis thaliana BOR1 (AtBOR1) variants based (i) on known disease causing mutations of hAE1 (S466R, A500R) and (ii) a loss of function mutation (D311A) identified in the yeast BOR protein, ScBOR1p. The AtBOR1 variants express in yeast and localise to the plasma membrane, although both S466R and A500R exhibit lower expression than the WT AtBOR1 and D311A. The D311A, S466R and A500R mutations result in a loss of borate efflux activity in a yeast bor1p knockout strain. A. thaliana plants containing these three individual mutations exhibit substantially decreased growth phenotypes in soil under conditions of low boron. These data confirm an important role for D311 in the function of the protein and show that mutations equivalent to disease-causing mutations in hAE1 have major effects in AtBOR1. We also obtained a low resolution cryo-EM structure of a BOR protein from Oryza sativa, OsBOR3, lacking the 30 C-terminal amino acid residues. This structure confirms the gate and core domain organisation previously observed for related proteins, and is strongly suggestive of an inward facing conformation.
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DOI：
10.1111/nph.13596
发表时间：
2016-01
期刊：
The New phytologist
影响因子：
--
作者：
Chormova D;Fry SC
通讯作者：
Fry SC
影响因子：
16.6
作者：
Huynh KW;Jiang J;Abuladze N;Tsirulnikov K;Kao L;Shao X;Newman D;Azimov R;Pushkin A;Zhou ZH;Kurtz I
通讯作者：
Kurtz I
影响因子：
14.8
作者：
Drew, David;Newstead, Simon;Iwata, So
通讯作者：
Iwata, So
影响因子：
6.9
作者：
Camacho-Cristóbal JJ;Martín-Rejano EM;Herrera-Rodríguez MB;Navarro-Gochicoa MT;Rexach J;González-Fontes A
通讯作者：
González-Fontes A
影响因子：
6.4
作者：
Li, CJ;Pfeffer, H;Bangerth, F
通讯作者：
Bangerth, F