Institut Jean-Pierre Bourgin, Unité Mixte de Recherche 1318 INRA-AgroParisTech, INRA Centre de Versailles-Grignon, Route de Saint-Cyr (RD10), Versailles Cedex, France.
Plant J. 2010 Oct;64(2):291-303. doi: 10.1111/j.1365-313X.2010.04332.x. Epub 2010 Sep 16.
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.
PII 蛋白是中心代谢和能量水平的整合者。在拟南芥中,细胞能量和碳水平的变构感应改变了 PII 与靶酶(如 N-乙酰-l-谷氨酸激酶和异源乙酰辅酶 A 羧化酶)相互作用的能力,从而调节这些质体 ATP 和碳消耗酶的生物学活性。定量逆转录聚合酶链反应方法显示,在早期种子成熟过程中,编码 PII 的 AtGLB1 基因(At4g01900)的表达增加了三倍。AtGLB1 启动子的活性与这种模式一致。一组互补的分子和遗传分析表明,WRINKLED1,一种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表明,PII 蛋白水平忠实地反映了 AtGLB1 mRNA 的积累。在亚细胞水平上,在成熟胚胎的质体中观察到 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。在突变体种子中观察到脂肪酸产量的短暂增加,此时在野生型种子中发现 PII 蛋白含量达到最大值。此外,在 pII 种子中测量到脂肪酸组成的微小但统计学上显著的修饰,这些种子表现出修饰(延长、去饱和)脂肪酸种类的减少。结果表明 PII 在脂肪酸生物合成和分配的精细调节中起作用。