Lin Ying-Chen, Liu Yu-Chi, Nakamura Yuki
Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan
Plant Cell. 2015 May;27(5):1497-511. doi: 10.1105/tpc.15.00207. Epub 2015 May 12.
Phospholipids are highly conserved and essential components of biological membranes. The major phospholipids, phosphatidylethanolamine and phosphatidylcholine (PtdCho), are synthesized by the transfer of the phosphoethanolamine or phosphocholine polar head group, respectively, to the diacylglycerol backbone. The metabolism of the polar head group characterizing each phospholipid class is poorly understood; thus, the biosynthetic pathway of major phospholipids remains elusive in Arabidopsis thaliana. The choline/ethanolamine kinase (CEK) family catalyzes the initial steps of phospholipid biosynthesis. Here, we analyzed the function of the four CEK family members present in Arabidopsis. Knocking out of CEK4 resulted in defective embryo development, which was complemented by transformation of genomic CEK4. Reciprocal genetic crossing suggested that CEK4 knockout causes embryonic lethality, and microscopy analysis of the aborted embryos revealed developmental arrest after the heart stage, with no defect being found in the pollen. CEK4 is preferentially expressed in the vasculature, organ boundaries, and mature embryos, and CEK4 was mainly localized to the plasma membrane. Overexpression of CEK4 in wild-type Arabidopsis increased the levels of PtdCho in seedlings and mature siliques and of major membrane lipids in seedlings and triacylglycerol in mature siliques. CEK4 may be the plasma membrane-localized isoform of the CEK family involved in the rate-limiting step of PtdCho biosynthesis and appears to be required for embryo development in Arabidopsis.
磷脂是生物膜中高度保守且必不可少的成分。主要的磷脂,磷脂酰乙醇胺和磷脂酰胆碱(PtdCho),分别通过将磷酸乙醇胺或磷酸胆碱极性头部基团转移到二酰基甘油主链上来合成。对于表征每种磷脂类别的极性头部基团的代谢了解甚少;因此,拟南芥中主要磷脂的生物合成途径仍然不清楚。胆碱/乙醇胺激酶(CEK)家族催化磷脂生物合成的起始步骤。在这里,我们分析了拟南芥中存在的四个CEK家族成员的功能。敲除CEK4导致胚胎发育缺陷,而基因组CEK4的转化可弥补这一缺陷。正反交遗传实验表明,CEK4敲除会导致胚胎致死,对流产胚胎的显微镜分析显示,心脏期后发育停滞,在花粉中未发现缺陷。CEK4在维管组织、器官边界和成熟胚胎中优先表达,并且CEK4主要定位于质膜。在野生型拟南芥中过表达CEK4会增加幼苗和成熟角果中PtdCho的水平,以及幼苗中主要膜脂和成熟角果中三酰甘油的水平。CEK4可能是CEK家族中定位于质膜的异构体,参与PtdCho生物合成的限速步骤,并且似乎是拟南芥胚胎发育所必需的。