Benenati Gaspare, Penkov Sider, Müller-Reichert Thomas, Entchev Eugeni V, Kurzchalia Teymuras V
Max-Planck-Institute for Molecular Cell Biology and Genetics, Dresden, Germany.
Mech Dev. 2009 May-Jun;126(5-6):382-93. doi: 10.1016/j.mod.2009.02.001. Epub 2009 Feb 11.
The role of lipids in the process of embryonic development of Caenorhabditis elegans is still poorly understood. Cytochrome P450s, a class of lipid-modifying enzymes, are good candidates to be involved in the production or degradation of lipids essential for development. We investigated two highly similar cytochrome P450s in C. elegans, cyp-31A2 and cyp-31A3, that are homologs of the gene responsible for Bietti crystalline corneoretinal dystrophy in humans. Depletion of both cytochromes either by RNAi or using a double deletion mutant, led to the failure of establishing the correct polarity of the embryo and to complete the extrusion of the polar bodies during meiosis. In addition, the egg became osmotic sensitive and permeable to dyes. The phenotype of cyp-31A2 or cyp-31A3 is very similar to a class of mutants that have polarization and osmotic defects (POD), thus the genes were renamed to pod-7 and pod-8, respectively. Electron microscopic analysis demonstrated that the activity of pod-7/pod-8 is crucial for the proper assembly of the eggshell and, in particular, for the production of its lipid-rich layer. Using a complementation with lipid extracts, we show that POD-7/POD-8 function together with a NADPH cytochrome P450 reductase, coded by emb-8, and are involved in the production of lipid(s) required for eggshell formation.
脂质在秀丽隐杆线虫胚胎发育过程中的作用仍知之甚少。细胞色素P450s是一类脂质修饰酶,是参与发育所需脂质产生或降解的良好候选者。我们研究了秀丽隐杆线虫中两种高度相似的细胞色素P450s,即cyp-31A2和cyp-31A3,它们是人类中导致比埃蒂结晶性角膜视网膜营养不良的基因的同源物。通过RNA干扰或使用双缺失突变体使这两种细胞色素缺失,会导致胚胎无法建立正确的极性,并在减数分裂期间无法完成极体的挤出。此外,卵变得对渗透压敏感且对染料具有渗透性。cyp-31A2或cyp-31A3的表型与一类具有极化和渗透缺陷(POD)的突变体非常相似,因此这两个基因分别被重新命名为pod-7和pod-8。电子显微镜分析表明,pod-7/pod-8的活性对于卵壳的正确组装至关重要,特别是对于其富含脂质层的产生。通过与脂质提取物互补,我们表明POD-7/POD-8与由emb-8编码的NADPH细胞色素P450还原酶共同发挥作用,并参与卵壳形成所需脂质的产生。