Klahre U, Noguchi T, Fujioka S, Takatsuto S, Yokota T, Nomura T, Yoshida S, Chua N H
Laboratory of Plant Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, New York 10021-6399, USA.
Plant Cell. 1998 Oct;10(10):1677-90. doi: 10.1105/tpc.10.10.1677.
We have identified the function of the Arabidopsis DIMINUTO/DWARF1 (DIM/DWF1) gene by analyzing the dim mutant, a severe dwarf with greatly reduced fertility. Both the mutant phenotype and gene expression could be rescued by the addition of exogenous brassinolide. Analysis of endogenous sterols demonstrated that dim accumulates 24-methylenecholesterol but is deficient in campesterol, an early precursor of brassinolide. In addition, we show that dim is deficient in brassinosteroids as well. Feeding experiments using deuterium-labeled 24-methylenecholesterol and 24-methyldesmosterol confirmed that DIM/DWF1 is involved in both the isomerization and reduction of the Delta24(28) bond. This conversion is not required in cholesterol biosynthesis in animals but is a key step in the biosynthesis of plant sterols. Transient expression of a green fluorescent protein-DIM/DWF1 fusion protein and biochemical experiments showed that DIM/DWF1 is an integral membrane protein that most probably is associated with the endoplasmic reticulum.
我们通过分析拟南芥DIMINUTO/DWARF1(DIM/DWF1)基因的dim突变体(一种严重矮化且育性大幅降低的突变体)确定了该基因的功能。添加外源油菜素内酯可挽救突变体表型和基因表达。对内源甾醇的分析表明,dim积累24-亚甲基胆固醇,但缺乏油菜素内酯的早期前体菜油甾醇。此外,我们还表明dim也缺乏油菜素甾醇。使用氘标记的24-亚甲基胆固醇和24-甲基去氢胆甾醇进行的饲喂实验证实,DIM/DWF1参与Δ24(28)键的异构化和还原。这种转化在动物胆固醇生物合成中并非必需,但却是植物甾醇生物合成中的关键步骤。绿色荧光蛋白-DIM/DWF1融合蛋白的瞬时表达和生化实验表明,DIM/DWF1是一种整合膜蛋白,极有可能与内质网相关。