Sperling R, Spann P, Offen D, Sperling J
Proc Natl Acad Sci U S A. 1986 Sep;83(18):6721-5. doi: 10.1073/pnas.83.18.6721.
Nuclear ribonucleoprotein (RNP) complexes that contain intact transcripts of the amplified gene for CAD, the multifunctional protein that initiates UMP synthesis in Syrian hamster cells, have been released from nuclei of Syrian hamster cells as large particulate structures that sediment at the 200S region in a sucrose gradient. By the technique of RNA hybridization, we have shown that U1, U2, and U6 small nuclear RNAs (snRNAs) cosediment with the large RNP particles in the sucrose gradients. Autoimmune sera from systemic lupus erythematosus and mixed connective tissue disease patients, characterized as anti-(U1)RNP, have further been shown to immunoprecipitate CAD RNA along with U1 and U2 snRNAs from the fractionated nuclear 200S RNP particles. We conclude that U1, U2, and U6 snRNPs are integral constituents of the 200S RNP particles. The requirement of snRNPs for RNA processing that evidently occurs on RNP particles has been recently demonstrated. Our results thus suggest that the 200S RNPs are structurally and functionally close to the native particles on which RNA processing occurs.
核核糖核蛋白(RNP)复合物含有叙利亚仓鼠细胞中CAD扩增基因的完整转录本,CAD是启动叙利亚仓鼠细胞UMP合成的多功能蛋白,该复合物已从叙利亚仓鼠细胞核中释放出来,成为在蔗糖梯度中200S区域沉降的大颗粒结构。通过RNA杂交技术,我们已经表明U1、U2和U6小核RNA(snRNA)与蔗糖梯度中的大RNP颗粒共沉降。来自系统性红斑狼疮和混合性结缔组织病患者的自身免疫血清,其特征为抗(U1)RNP,进一步被证明可从分级分离的核200S RNP颗粒中与U1和U2 snRNA一起免疫沉淀CAD RNA。我们得出结论,U1、U2和U6 snRNP是200S RNP颗粒的组成成分。最近已经证明了snRNP对显然在RNP颗粒上发生的RNA加工的需求。因此,我们的结果表明,200S RNP在结构和功能上与发生RNA加工的天然颗粒接近。