Hurt E C
EMBL, Heidelberg, FRG.
EMBO J. 1988 Dec 20;7(13):4323-34. doi: 10.1002/j.1460-2075.1988.tb03331.x.
In order to study the role of nucleoskeletal components for nuclear and cell division in the budding yeast Saccharomyces cerevisiae, we have employed a combined biochemical/genetic approach. We have identified a peripheral nuclear protein which appears to be located both at the nuclear membrane and the spindle pole body. The gene has been cloned and subsequently shown to be essential for cell growth. The DNA sequence of the gene has been determined. As deduced from the nucleotide sequence, the gene potentially codes for a novel 86 kd protein with a highly repetitive and conserved nine amino acid sequence motive in the middle part of the protein. The flanking amino- and carboxy-terminal regions have similarities to intermediate filaments and calcium binding proteins, respectively. It appears that the 86 kd protein is a regulated nucleoskeletal-like protein (NSP1) involved in the process of nuclear and/or cell division. The affinity-purified antibody against the yeast NSP1 protein stained the nucleus and centrosomes of mammalian MDCK (Madin Darby canine kidney) cells in indirect immunofluorescence.
为了研究核骨架成分在出芽酵母酿酒酵母的细胞核和细胞分裂中的作用,我们采用了生化/遗传相结合的方法。我们鉴定出一种位于细胞核周边的蛋白质,它似乎同时定位于核膜和纺锤极体。该基因已被克隆,随后显示其对细胞生长至关重要。该基因的DNA序列已被测定。从核苷酸序列推断,该基因可能编码一种新的86千道尔顿的蛋白质,该蛋白质在其中部具有高度重复且保守的九氨基酸序列基序。其侧翼的氨基末端和羧基末端区域分别与中间丝和钙结合蛋白相似。看来,这种86千道尔顿的蛋白质是一种受调控的核骨架样蛋白(NSP1),参与细胞核和/或细胞分裂过程。抗酵母NSP1蛋白的亲和纯化抗体在间接免疫荧光中对哺乳动物MDCK(马-达二氏犬肾)细胞的细胞核和中心体进行了染色。