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PII is induced by WRINKLED1 and fine-tunes fatty acid composition in seeds of Arabidopsis thaliana

机译:PII由WRINKLED1诱导并微调拟南芥种子中的脂肪酸组成

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摘要

P>The PII protein is an integrator of central metabolism and energy levels. In Arabidopsis, allosteric sensing of cellular energy and carbon levels alters the ability of PII to interact with target enzymes such as N-acetyl-l-glutamate kinase and heteromeric acetyl-coenzyme A carboxylase, thereby modulating the biological activity of these plastidial ATP- and carbon-consuming enzymes. A quantitative reverse transcriptase-polymerase chain reaction approach revealed a threefold induction of the AtGLB1 gene (At4g01900) encoding PII during early seed maturation. The activity of the AtGLB1 promoter was consistent with this pattern. A complementary set of molecular and genetic analyses showed that WRINKLED1, a transcription factor known to induce glycolytic and fatty acid biosynthetic genes at the onset of seed maturation, directly controls AtGLB1 expression. Immunoblot analyses and immunolocalization experiments using anti-PII antibodies established that PII protein levels faithfully reflected AtGLB1 mRNA accumulation. At the subcellular level, PII was observed in plastids of maturing embryos. To further investigate the function of PII in seeds, comprehensive functional analyses of two pII mutant alleles were carried out. A transient increase in fatty acid production was observed in mutant seeds at a time when PII protein content was found to be maximal in wild-type seeds. Moreover, minor though statistically significant modifications of the fatty acid composition were measured in pII seeds, which exhibited decreased amounts of modified (elongated, desaturated) fatty acid species. The results obtained outline a role for PII in the fine tuning of fatty acid biosynthesis and partitioning in seeds.
机译:P> PII蛋白是中枢代谢和能量水平的整合剂。在拟南芥中,对细胞能量和碳水平的变构感测会改变PII与目标酶(例如N-乙酰基-1-谷氨酸激酶和异聚乙酰基辅酶A羧化酶)相互作用的能力,从而调节这些质体ATP-和耗碳酶。定量逆转录酶-聚合酶链反应方法揭示了在种子早期成熟过程中编码PII的AtGLB1基因(At4g01900)的三倍诱导。 AtGLB1启动子的活性与此模式一致。分子和遗传分析的补充集表明,WRINKLED1是已知在种子成熟开始时诱导糖酵解和脂肪酸生物合成基因的转录因子,直接控制AtGLB1的表达。使用抗PII抗体的免疫印迹分析和免疫定位实验确定,PII蛋白水平忠实地反映了AtGLB1 mRNA的积累。在亚细胞水平上,在成熟胚的质体中观察到PII。为了进一步研究PII在种子中的功能,对两个pII突变等位基因进行了全面的功能分析。当发现野生型种子中PII蛋白含量最大时,突变种子中脂肪酸的产量会短暂增加。此外,在pII种子中测定了脂肪酸组成的微小但统计学上显着的修饰,其显示出减少的修饰(伸长,去饱和)脂肪酸种类的量。获得的结果概述了PII在脂肪酸生物合成的精细调节和种子中分配方面的作用。

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