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A Chloroplastic UDP-Glucose Pyrophosphorylase from Arabidopsis Is the Committed Enzyme for the First Step of Sulfolipid Biosynthesis

机译:来自拟南芥的叶绿体UDP-葡萄糖焦磷酸化酶是糖脂生物合成第一步的必不可少的酶。

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摘要

Plants synthesize a sulfur-containing lipid, sulfoquinovosyldiacylglycerol, which is one of three nonphosphorus glycerolipids that provide the bulk of the structural lipids in photosynthetic membranes. Here, the identification of a novel gene, UDP-glucose pyrophosphorylase3 (UGP3), required for sulfolipid biosynthesis is described. Transcriptome coexpression analysis demonstrated highly correlated expression of UGP3 with known genes for sulfolipid biosynthesis in Arabidopsis thaliana. Liquid chromatography-mass spectrometry analysis of leaf lipids in two Arabidopsis ugp3 mutants revealed that no sulfolipid was accumulated in these mutants, indicating the participation of UGP3 in sulfolipid biosynthesis. From the deduced amino acid sequence, UGP3 was presumed to be a UDP-glucose pyrophosphorylase (UGPase) involved in the generation of UDP-glucose, serving as the precursor of the polar head of sulfolipid. Recombinant UGP3 was able to catalyze the formation of UDP-glucose from glucose-1-phosphate and UTP. A transient assay using fluorescence fusion proteins and UGPase activity in isolated chloroplasts indicated chloroplastic localization of UGP3. The transcription level of UGP3 was increased by phosphate starvation. A comparative genomics study on UGP3 homologs across different plant species suggested the structural and functional conservation of the proteins and, thus, a committing role for UGP3 in sulfolipid synthesis.
机译:植物合成了一种含硫脂质磺基喹喔基二酰基甘油,这是三种非磷甘油脂之一,可在光合膜中提供大部分结构脂质。在此,描述了脂脂质生物合成所需的新基因UDP-葡萄糖焦磷酸化酶3(UGP3)的鉴定。转录组共表达分析表明拟南芥中UGP3的表达与硫脂类生物合成的已知基因高度相关。两种拟南芥ugp3突变体中叶脂的液相色谱-质谱分析表明,这些突变体中未积累硫脂,表明UGP3参与了硫脂的生物合成。根据推导的氨基酸序列,认为UGP3是涉及UDP-葡萄糖的生成的UDP-葡萄糖焦磷酸化酶(UGPase),它是硫脂极性极的前体。重组UGP3能够催化由1-磷酸葡萄糖和UTP形成UDP-葡萄糖。在分离的叶绿体中使用荧光融合蛋白和UGPase活性进行的瞬时测定表明UGP3的叶绿体定位。 UGP3的转录水平由于磷酸盐饥饿而增加。一项关于跨不同植物物种的UGP3同源物的比较基因组学研究表明,蛋白质的结构和功能保守,因此,UGP3在硫脂合成中具有重要作用。

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