Harper Nicola C, Al-Greene Nicole T, Basrai Munira A, Belanger Kenneth D
Department of Biology, Colgate University, NY 13346, USA.
Curr Genet. 2008 Feb;53(2):95-105. doi: 10.1007/s00294-007-0168-4. Epub 2007 Dec 5.
Nuclear pore complexes (NPCs) are embedded in the nuclear envelope of eukaryotic cells and function to regulate passage of macromolecules in and out of the nucleus. Nup1 is one of 30 nucleoporins comprising the NPC of the yeast Saccharomyces cerevisiae and is located on the nucleoplasmic face of the NPC where it plays a role in mRNA export and protein transport. In order to further characterize the function of Nup1 we used a genetic approach to identify mutations that are synthetically lethal in combination with a deletion of NUP1 (nup1Delta). We have identified one such nup1 lethal mutant (nle6) as a temperature sensitive allele of nud1. NUD1 encodes a component of the yeast spindle pole body (SPB) and acts as scaffolding for the mitotic exit network (MEN). We observe that nle6/nud1 mutant cells have a normal distribution of NPCs within the nuclear envelope and exhibit normal rates of nuclear protein import at both the permissive and restrictive temperatures. nup1Delta also exhibits synthetic lethality with bub2Delta and bfa1Delta, both of which encode proteins that colocalize with Nud1 at spindle pole bodies and function in the mitotic exit network. However, we do not observe genetic interactions among nle6/nud1, bub2Delta, or bfa1Delta and mutations in the nucleoporin encoding genes NUP60 or NUP170, nor is nup1Delta synthetically lethal with the absence of components downstream in the mitotic exit network, including Lte1, Swi5, and Dbf2. Our results suggest a novel functional connection between Nup1 and proteins comprising both the spindle pole body and early mitotic exit network.
核孔复合体(NPCs)嵌入真核细胞的核膜中,其功能是调节大分子进出细胞核。Nup1是构成酿酒酵母NPC的30种核孔蛋白之一,位于NPC的核质面上,在mRNA输出和蛋白质运输中发挥作用。为了进一步表征Nup1的功能,我们采用遗传学方法来鉴定与NUP1缺失(nup1Delta)组合时具有合成致死性的突变。我们已鉴定出一个这样的nup1致死突变体(nle6),它是nud1的温度敏感等位基因。NUD1编码酵母纺锤体极体(SPB)的一个组分,并作为有丝分裂退出网络(MEN)的支架。我们观察到,nle6/nud1突变细胞在核膜内NPC分布正常,并且在允许温度和限制温度下均表现出正常的核蛋白输入速率。nup1Delta与bub2Delta和bfa1Delta也表现出合成致死性,这两者均编码与Nud1在纺锤体极体共定位并在有丝分裂退出网络中起作用的蛋白质。然而,我们未观察到nle6/nud1、bub2Delta或bfa1Delta与核孔蛋白编码基因NUP60或NUP170中的突变之间存在遗传相互作用,nup1Delta在缺少有丝分裂退出网络下游组分(包括Lte1、Swi5和Dbf2)时也不具有合成致死性。我们的结果表明Nup1与构成纺锤体极体和早期有丝分裂退出网络的蛋白质之间存在一种新的功能联系。