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Arabidopsis ABCB21 is a Facultative Auxin Importer/Exporter Regulated by Cytoplasmic Auxin Concentration

机译:拟南芥ABCB21是细胞质生长素浓度调节的兼性生长素输入/输出

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摘要

The phytohormone auxin is critical for plant growth and many developmental processes. Members of the P-glycoprotein (PGP/ABCB) subfamily of ATP-binding cassette (ABC) transporters have been shown to function in the polar movement of auxin by transporting auxin over the plasma membrane in both monocots and dicots. Here, we characterize a new Arabidopsis member of the ABCB subfamily, ABCB21/PGP21, a close homolog of ABCB4, for which conflicting transport directionalities have been reported. ABCB21 is strongly expressed in the abaxial side of cotyledons and in junctions of lateral organs in the aerial part, whereas in roots it is specifically expressed in pericycle cells. Membrane fractionation by sucrose density gradient centrifugation followed by Western blot showed that ABCB21 is a plasma membrane-localized ABC transporter. A transport assay with Arabidopsis protoplasts suggested that ABCB21 was involved in IAA transport in an outward direction, while naphthalene acetic acid (NAA) was a less preferable substrate for ABCB21. Further functional analysis of ABCB21 using yeast import and export assays showed that ABCB21 mediates the 1-N-naphthylphthalamic acid (NPA)-sensitive translocation of auxin in an inward direction when the cytoplasmic IAA concentration is low, whereas this transporter mediates outward transport under high internal IAA. An increase in the cytoplasmic IAA concentration by pre-loading of IAA into yeast cells abolished the IAA uptake activity by ABCB21 as well as ABCB4. These findings suggest that ABCB21 functions as a facultative importer/exporter controlling auxin concentrations in plant cells
机译:植物激素生长素对于植物生长和许多发育过程至关重要。 ATP结合盒(ABC)转运蛋白的P-糖蛋白(PGP / ABCB)亚家族成员已显示出通过在单子叶植物和双子叶植物中通过在细胞膜上运输生长素来在生长素的极性运动中起作用。在这里,我们表征了ABCB亚家族的一个新的拟南芥成员ABCB21 / PGP21,它是ABCB4的近亲同源物,其运输方向相互矛盾。 ABCB21在子叶的背面和空中部分的横向器官的交界处强烈表达,而在根中则在周周细胞中特异性表达。蔗糖密度梯度离心分离膜,然后进行Western印迹显示,ABCB21是质膜定位的ABC转运蛋白。拟南芥原生质体的转运分析表明,ABCB21参与了IAA向外的转运,而萘乙酸(NAA)是ABCB21的次佳底物。使用酵母进出口检测方法对ABCB21进行的进一步功能分析表明,当细胞质IAA浓度较低时,ABCB21会向内介导生长素的1-N-萘基邻苯二甲酸(NPA)敏感转运,而在高浓度下该转运蛋白会介导向外转运内部IAA。通过将IAA预装到酵母细胞中,细胞质IAA浓度的增加消除了ABCB21和ABCB4对IAA的吸收活性。这些发现表明,ABCB21充当控制植物细胞中生长素浓度的兼职进出口者。

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