对绿色荧光蛋白(GFP)融合文库进行可视化筛选,鉴定出一种新型核被膜蛋白。
A visual screen of a GFP-fusion library identifies a new type of nuclear envelope membrane protein.
作者信息
Rolls M M, Stein P A, Taylor S S, Ha E, McKeon F, Rapoport T A
机构信息
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
出版信息
J Cell Biol. 1999 Jul 12;146(1):29-44. doi: 10.1083/jcb.146.1.29.
The nuclear envelope (NE) is a distinct subdomain of the ER, but few membrane components have been described that are specific to it. We performed a visual screen in tissue culture cells to identify proteins targeted to the NE. This approach does not require assumptions about the nature of the association with the NE or the physical separation of NE and ER. We confirmed that screening a library of fusions to the green fluorescent protein can be used to identify proteins targeted to various subcompartments of mammalian cells, including the NE. With this approach, we identified a new NE membrane protein, named nurim. Nurim is a multispanning membrane protein without large hydrophilic domains that is very tightly associated with the nucleus. Unlike the known NE membrane proteins, it is neither associated with nuclear pores, nor targeted like lamin-associated membrane proteins. Thus, nurim is a new type of NE membrane protein that is localized to the NE by a distinct mechanism.
核膜(NE)是内质网(ER)的一个独特亚结构域,但针对其的特异性膜成分却鲜有报道。我们在组织培养细胞中进行了一项可视化筛选,以鉴定靶向核膜的蛋白质。这种方法无需对与核膜的关联性质或核膜与内质网的物理分离进行假设。我们证实,筛选与绿色荧光蛋白的融合文库可用于鉴定靶向哺乳动物细胞各种亚区室(包括核膜)的蛋白质。通过这种方法,我们鉴定出一种新的核膜蛋白,命名为nurim。Nurim是一种多跨膜蛋白,没有大的亲水区,与细胞核紧密相关。与已知的核膜蛋白不同,它既不与核孔相关,也不像核纤层相关膜蛋白那样靶向定位。因此,nurim是一种新型的核膜蛋白,通过独特的机制定位于核膜。