Identification of phospholipid translocase and its role in cell polarity formation.

磷脂转位酶的鉴定及其在细胞极性形成中的作用。

• 批准号: 12480220
• 负责人: UMEDA Masato
• 金额: $ 8.9万
• 依托单位: Tokyo Metropolitan Organization for Medical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480220/
• 关键词: phospholipid; cytoskeleton; lipid bilayer; membrane; cell polarity; actin; yeast; phosphatidylethanolamine; 細胞骨格; 細胞膜; 細胞分裂; 分裂溝; 神経細胞

It is well established that phospholipids in biological membranes are distributed asymmetrically between the inner and outer leaflets of the lipid bilayer. Although recent studies have shown that the transbilayer lipid asymmetry is generated and controlled by a family of specific lipid transport proteins, the physiological role of the lipid asymmetry remains largely unknown. Over the past decade, we have established a series of phospholipid-binding probes and mammalian cell mutants defective in phospholipid biosynthesis. These probes and mutants have provided useful tools to study the molecular motion and the cellular function of membrane phospholipids.In the final stage of cell division, cytokinesis constricts and then seals the plasma membrane between the two daughter cells. The constriction is powered by a contractile ring of actin, and scission involves a fusion or rearrangement of the lipid bilayer of the cell membrane. Using phospholipid-specific binding probes that we have gener … More ated over the past decade, we found that the lipid phosphatidylethanolamine (PE), which normally resides in the inner leaflet of the bilayer, was exposed onto the outer leaflet of the cleavage furrow. Subsequent analyses, using the phospholipid-binding probes and mutant cells defective in PE synthesis, have shown that this surface exposure of PE on the cleavage furrow is needed to coordinate the reorganization of the actin cytoskeleton and the plasma membrane during cytokinesis.To identify the molecules involved in the lipid-cytoskeleton coordination, we have isolated budding yeast mutants that have a defective in the transbilayer movement of phospholipids. A gene, designated as ROS3, was identified as a regulator of transbilayer relocation of PE on plasma membrane. ROS3 encodes a novel transmembrane protein present on plasma membrane. Disruption of ROS3 resulted in defects in morphology and formation of cortical actin patch. Overexpression of Ros3p led to multibud formation. Since Ros3p homologue is strongly expressed in some mammalian tissues such as brain and epithelial cells, Ros3p may play a general role in possible cross-talks between the cytoskeleton and membrane lipids that are required for proper cell division as well as cell polarity formation. Less

众所周知，生物膜中的磷脂在脂质双层的内外小叶之间不对称地分布。虽然最近的研究表明，跨双层脂质不对称性是由一个家庭的特定的脂质转运蛋白产生和控制，脂质不对称性的生理作用仍然在很大程度上未知。在过去的十年中，我们已经建立了一系列的磷脂结合探针和哺乳动物细胞突变体的磷脂生物合成缺陷。这些探针和突变体为研究膜磷脂的分子运动和细胞功能提供了有用的工具。在细胞分裂的最后阶段，胞质分裂收缩并密封两个子细胞之间的质膜。收缩是由肌动蛋白的收缩环提供动力，而断裂涉及细胞膜脂质双层的融合或重排。利用我们现有的磷脂特异性结合探针， ...更多信息 在过去的十年中，我们发现，脂质磷脂酰乙醇胺（PE），这通常驻留在双层的内小叶，是暴露在分裂沟的外小叶。随后的分析，使用磷脂结合探针和突变体细胞在PE合成缺陷，已经表明，这种表面暴露的PE上的分裂沟是需要协调重组的肌动蛋白细胞骨架和质膜在胞质分裂。为了确定参与的脂质细胞骨架协调的分子，我们已经分离出芽殖酵母突变体，有缺陷的transbilayer运动的磷脂。一个被命名为ROS 3的基因被鉴定为PE在质膜上跨双层迁移的调节因子。ROS3编码一种存在于质膜上的新型跨膜蛋白。ROS3的破坏导致皮层肌动蛋白斑块的形态和形成缺陷。Ros3p的过表达导致多芽形成。由于Ros3p同源物在一些哺乳动物组织如脑和上皮细胞中强烈表达，因此Ros3p可能在细胞骨架和膜脂质之间的可能的交联中发挥一般作用，所述交联是适当的细胞分裂以及细胞极性形成所需的。少

项目成果

Kato, U., Emoto, K., Fredriksson, C., Nakamura, H., Ohta, A., Kobayashi, T., Murakami-Murofushi, K., Kobayashi, T., Umeda. M.: "A novel membrane protein, Ros3p, is required for phospholipid translocation across the plasma membrane in Saccharomyces cerevis

加藤，U.，江本，K.，弗雷德里克森，C.，中村，H.，太田，A.，小林，T.，村上室伏，K.，小林，T.，梅田。

Yokogawa, T., Nagata, S., Nishio, Y., Tsutsumi, T., Ihara, S., Shirai, R., Morita, K., Umeda, M., Shirai, Y., Saito, N., Fukui, Y.: "Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface : Use of immunostaining techniq

横河，T.，永田，S.，西尾，Y.，堤，T.，伊原，S.，白井，R.，森田，K.，梅田，M.，白井，Y.，斋藤，N.，

Nakai, Y., Sakurai, Y., Yamaji, A., Umeda, M., Asou, H., Uyemura, K., Itoh, K.: "Lysenin-Sphingomyelin binding at the cell surface of oligodendrocyte linage cells increases during differentiation in vitro"J. Neurosci. Res.. 62. 521-529 (2000)

Nakai, Y.、Sakurai, Y.、Yamaji, A.、Umeda, M.、Asou, H.、Uyemura, K.、Itoh, K.：“少突胶质细胞系细胞表面的赖森素-鞘磷脂结合在

Harada, A. et al.: "Nadrin, an novel neuron-specific GTPase activating protein involved in regulated exocytosis"J.Biol.Chem.. 275. 36885-36891 (2000)

Harada, A. 等人：“Nadrin，一种参与调节胞吐作用的新型神经元特异性 GTP 酶激活蛋白”J.Biol.Chem.. 275. 36885-36891 (2000)

Harada, A., Furuta, B., Takeuchi, K., Itakura, M., Takahashi, M., Umeda, M.: "Nadrin, a novel neuron-specific GTPase activating protein, implicated in regulated exocytosis"Neurosci. Res.. 24. S89 (2001)

Harada, A.、Furuta, B.、Takeuchi, K.、Itakura, M.、Takahashi, M.、Umeda, M.：“Nadrin，一种新型神经元特异性 GTP 酶激活蛋白，与调节的胞吐作用有关”Neurosci。