Kondo K, Inouye M
Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey at Rutgers, Piscataway 08854-5635.
J Biol Chem. 1992 Aug 15;267(23):16252-8.
We have identified a yeast gene encoding a protein structurally similar to mammalian nucleolin. The gene was previously cloned as a cold shock-inducible gene and found to be identical to yeast NSR1 gene, which encodes a protein that has been reported to bind sequences required for nuclear localization of protein. The carboxyl-terminal half of NSR1, consisting of two tandemly repeated putative RNA-binding domains and a glycine/arginine-rich domain, has 37% amino acid sequence identity with the same part of mammalian nucleolin, while no sequence similarities are found between their amino-terminal regions. Although a null mutation of the NSR1 gene was not lethal, it caused a severe defect on growth. Pulse-labeling analysis revealed that the nsr1 strain had reduced levels of 18 S rRNA and accumulated 35 S pre-rRNA compared with the wild-type strain. The level of 25 S rRNA was also slightly reduced in the nsr1 strain. Pulse-chase labeling experiments showed slow processing of 35 S pre-rRNA and impaired methylation of 18 S rRNA. The ratio of 40 S to 60 S ribosomal subunits in the nsr1 strain is significantly reduced and is consistent with impaired synthesis of 18 S rRNA. The results indicate that NSR1 is involved in pre-rRNA processing and ribosome biosynthesis in yeast.
我们已鉴定出一个编码与哺乳动物核仁素结构相似蛋白质的酵母基因。该基因先前作为冷休克诱导基因被克隆,发现与酵母NSR1基因相同,NSR1基因编码的蛋白质据报道可结合蛋白质核定位所需的序列。NSR1的羧基末端一半由两个串联重复的假定RNA结合结构域和一个富含甘氨酸/精氨酸的结构域组成,与哺乳动物核仁素的相同部分具有37%的氨基酸序列同一性,而它们的氨基末端区域之间未发现序列相似性。尽管NSR1基因的无效突变并不致命,但它导致了严重的生长缺陷。脉冲标记分析显示,与野生型菌株相比,nsr1菌株中18 S rRNA水平降低,35 S前体rRNA积累。nsr1菌株中25 S rRNA水平也略有降低。脉冲追踪标记实验表明35 S前体rRNA加工缓慢,18 S rRNA甲基化受损。nsr1菌株中40 S与60 S核糖体亚基的比例显著降低,这与18 S rRNA合成受损一致。结果表明,NSR1参与酵母中的前体rRNA加工和核糖体生物合成。