Pecot Matt Yasuo, Malhotra Vivek
Cell and Developmental Biology Department, University of California San Diego, La Jolla, CA 92093, USA.
Cell. 2004 Jan 9;116(1):99-107. doi: 10.1016/s0092-8674(03)01068-7.
What happens to organelles during mitosis, and how they are apportioned to each of the daughter cells, is not completely clear. We have devised a procedure to address whether Golgi membranes fuse with the Endoplasmic Reticulum (ER) during mitosis via the detection of interactions between ER and Golgi proteins. This procedure involves coexpressing an FKBP-tagged Golgi enzyme with an ER-retained protein fused to FRAP in COS cells. Since treatment with rapamycin induces a tight association between FKBP and FRAP, one would expect rapamycin to trap the FKBP-fused Golgi protein in the ER if it ever visits the ER during mitosis. However, after the doubly transfected cells progress through mitosis in the presence of rapamycin, we find the Golgi protein in the newly formed Golgi stacks and not in the ER. Based on these results, we conclude that Golgi membranes remain separate from the ER during mitosis in mammalian cells.
在有丝分裂过程中细胞器会发生什么，以及它们如何分配到每个子细胞中，目前尚不完全清楚。我们设计了一种方法，通过检测内质网（ER）和高尔基体蛋白之间的相互作用，来研究高尔基体膜在有丝分裂期间是否与内质网融合。该方法包括在COS细胞中共表达带有FKBP标签的高尔基体酶和与FRAP融合的内质网驻留蛋白。由于雷帕霉素处理会诱导FKBP和FRAP紧密结合，如果在有丝分裂期间FKBP融合的高尔基体蛋白进入内质网，人们会预期雷帕霉素会将其困在内质网中。然而，在双重转染的细胞在雷帕霉素存在的情况下经历有丝分裂后，我们发现高尔基体蛋白存在于新形成的高尔基体堆栈中，而不是内质网中。基于这些结果，我们得出结论，在哺乳动物细胞有丝分裂期间，高尔基体膜与内质网保持分离。
