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通过体内竞争试验揭示了酿酒酵母中的U1小核核糖核蛋白颗粒-蛋白质相互作用。

U1 small nuclear ribonucleoprotein particle-protein interactions are revealed in Saccharomyces cerevisiae by in vivo competition assays.

作者信息

Stutz F, Liao X C, Rosbash M

机构信息

Howard Hughes Medical Institute, Department of Biology, Brandeis University, Waltham, Massachusetts 02254.

出版信息

Mol Cell Biol. 1993 Apr;13(4):2126-33. doi: 10.1128/mcb.13.4.2126-2133.1993.

Abstract

Two highly conserved regions of the 586-nucleotide yeast (Saccharomyces cerevisiae) U1 small nuclear RNA (snRNA) can be mutated or deleted with little or no effect on growth rate: the universally conserved loop II (corresponding to the metazoan A loop) and the yeast core region (X. Liao, L. Kretzner, B. Séraphin, and M. Rosbash, Genes Dev. 4:1766-1774, 1990). To examine the contribution of these regions to U1 small nuclear ribonucleoprotein particle (snRNP) activity, a competitor U1 gene, encoding a nonfunctional U1 snRNA molecule, was introduced into a number of strains carrying a U1 snRNA gene with loop II or yeast core mutations. The presence of the nonfunctional U1 gene lowered the growth rate of these mutant strains but not wild-type strains, consistent with the notion that mutant U1 RNAs are less active than wild-type U1 snRNAs. A detailed analysis of the U1 snRNA levels and half-lives in a number of merodiploid strains suggests that these mutant U1 snRNAs interact with U1 snRNP proteins less well than do their wild-type counterparts. Competition for protein factors during snRNP assembly could account for a number of previous observations in both yeast and mammalian cells.

摘要

586个核苷酸的酵母(酿酒酵母)U1小核RNA(snRNA)的两个高度保守区域可以被突变或缺失,而对生长速率几乎没有影响:普遍保守的环II(对应于后生动物的A环)和酵母核心区域(X.廖,L.克雷茨纳,B.塞拉芬,和M.罗斯巴什,《基因与发育》4:1766 - 1774, 1990)。为了研究这些区域对U1小核糖核蛋白颗粒(snRNP)活性的贡献,将一个编码无功能U1 snRNA分子的竞争U1基因导入了一些携带具有环II或酵母核心突变的U1 snRNA基因的菌株中。无功能U1基因的存在降低了这些突变菌株的生长速率,但没有降低野生型菌株的生长速率,这与突变U1 RNA比野生型U1 snRNA活性更低的观点一致。对一些部分二倍体菌株中U1 snRNA水平和半衰期的详细分析表明,这些突变U1 snRNA与U1 snRNP蛋白的相互作用不如野生型U1 snRNA与U1 snRNP蛋白的相互作用好。snRNP组装过程中对蛋白质因子的竞争可以解释酵母和哺乳动物细胞中先前的一些观察结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f81/359533/ac58971cbecf/molcellb00016-0161-a.jpg

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