Holzinger Andreas, Lütz-Meindl Ursula
Institute of Plant Physiology, University of Salzburg, Austria.
Cell Biol Int. 2002;26(8):689-97. doi: 10.1006/cbir.2002.0920.
The unicellular green alga Micrasterias denticulata performs a two-directional postmitotic nuclear migration during development, a passive migration into the growing semicell, and a microtubule mediated backward migration towards the cell centre. The present study provides first evidence for force generation by motor proteins of the kinesin family in this process. The new kinesin specific inhibitor adociasulfate-2 causes abnormal nuclear displacement at 18 microM. AMP-PNP, a non hydrolyseable ATP analogue or the general ATPase inhibitors calyculin A and sodium orthovanadate also disturb nuclear migration. In addition kinesin-like proteins are detected by means of immunoblotting using antibodies against brain kinesin, plant derived antibodies to kinesin-like proteins and a calmodulin binding kinesin-like protein. Immunoelectron microscopy suggests a correlation of conventional kinesin-like proteins, but not of the calmodulin binding kinesin-like protein to the microtubule apparatus associated with the migrating nucleus.
单细胞绿藻齿缘微星藻在发育过程中进行双向有丝分裂后核迁移,一种是被动迁移到不断生长的半细胞中,另一种是微管介导的向细胞中心的反向迁移。本研究首次提供了在此过程中驱动蛋白家族的驱动蛋白产生力的证据。新型驱动蛋白特异性抑制剂阿多硫酸-2在18微摩尔浓度时会导致异常的核移位。AMP-PNP(一种不可水解的ATP类似物)或通用ATP酶抑制剂花萼海绵诱癌素A和原钒酸钠也会干扰核迁移。此外,利用抗脑驱动蛋白抗体、植物源抗驱动蛋白样蛋白抗体和一种钙调蛋白结合驱动蛋白样蛋白,通过免疫印迹法检测到了驱动蛋白样蛋白。免疫电子显微镜显示,传统驱动蛋白样蛋白与迁移核相关的微管装置存在相关性,但钙调蛋白结合驱动蛋白样蛋白与微管装置不存在相关性。