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首页> 外文期刊>Journal of Experimental Botany >Two solanesyl diphosphate synthases with different subcellular localizations and their respective physiological roles in Oryza sativa
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Two solanesyl diphosphate synthases with different subcellular localizations and their respective physiological roles in Oryza sativa

机译：两个不同亚细胞定位的茄基二磷酸茄酯合酶及其在水稻中的生理作用

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摘要

Long chain prenyl diphosphates are crucial biosynthetic precursors of ubiquinone (UQ) in many organisms, ranging from bacteria to humans, as well as precursors of plastoquinone in photosynthetic organisms. The cloning and characterization of two solanesyl diphosphate synthase genes, OsSPS1 and OsSPS2, in Oryza sativa is reported here. OsSPS1 was highly expressed in root tissue whereas OsSPS2 was found to be high in both leaves and roots. Enzymatic characterization using recombinant proteins showed that both OsSPS1 and OsSPS2 could produce solanesyl diphosphates as their final product, while OsSPS1 showed stronger activity than OsSPS2. However, an important biological difference was observed between the two genes: OsSPS1 complemented the yeast coq1 disruptant, which does not form UQ, whereas OsSPS2 only very weakly complemented the growth defect of the coq1 mutant. HPLC analyses showed that both OsSPS1 and OsSPS2 yeast transformants produced UQ9 instead of UQ6, which is the native yeast UQ. According to the complementation study, the UQ9 levels in OsSPS2 transformants were much lower than that of OsSPS1. Green fluorescent protein fusion analyses showed that OsSPS1 localized to mitochondria, while OsSPS2 localized to plastids. This suggests that OsSPS1 is involved in the supply of solanesyl diphosphate for ubiquinone-9 biosynthesis in mitochondria, whereas OsSPS2 is involved in providing solanesyl diphosphate for plastoquinone-9 formation. These findings indicate that O. sativa has a different mechanism for the supply of isoprenoid precursors in UQ biosynthesis from Arabidopsis thaliana, in which SPS1 provides a prenyl moiety for UQ9 at the endoplasmic reticulum.
机译：长链异戊二烯基二磷酸酯是从细菌到人类的许多生物中泛醌（UQ）的重要生物合成前体，以及光合生物中质体醌的前体。报道了水稻中两个茄基二磷酸二磷酸合酶基因OsSPS1和OsSPS2的克隆和鉴定。 OsSPS1在根组织中高表达，而OsSPS2在叶和根中均高表达。使用重组蛋白的酶促表征表明，OsSPS1和OsSPS2均可以产生茄基二磷酸酯作为最终产物，而OsSPS1则显示出比OsSPS2更强的活性。但是，在两个基因之间观察到了重要的生物学差异：OsSPS1补充了酵母coq1破坏物，后者不形成UQ，而OsSPS2仅非常弱地补充了coq1突变体的生长缺陷。 HPLC分析表明，OsSPS1和OsSPS2酵母转化子均产生UQ9而不是UQ6，后者是天然酵母UQ。根据互补研究，OsSPS2转化子中的UQ9水平远低于OsSPS1。绿色荧光蛋白融合分析表明，OsSPS1定位于线粒体，而OsSPS2定位于质体。这表明，OsSPS1参与线粒体中泛醌9生物合成的茄基二磷酸酯的供应，而OsSPS2参与质体醌9形成过程中的茄基二磷酸酯的提供。这些发现表明，苜蓿在拟南芥的UQ生物合成中提供类异戊二烯前体的机制有所不同，其中SPS1在内质网为UQ9提供异戊二烯基部分。

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