利用抗核糖核蛋白抗体检测与U1核糖核蛋白相关的不均一核核糖核蛋白中的β-珠蛋白信使核糖核酸前体。

Detection of beta-globin mRNA precursor in heterogeneous nuclear ribonucleoprotein associated with U1-RNP by using anti-RNP antibody.

作者信息

Wagatsuma M, Obinata M, Moroi Y, Hanaoka F, Yamada M

出版信息

J Biochem. 1985 Jun;97(6):1727-35. doi: 10.1093/oxfordjournals.jbchem.a135231.

DOI:10.1093/oxfordjournals.jbchem.a135231

Abstract

Heterogeneous nuclear RNA-ribonucleoprotein (hnRNP) fractions were isolated from Friend erythroleukemia cells and separated by 15-45% sucrose gradient centrifugation. The distribution of small nuclear RNAs (snRNAs) in hnRNP fractions indicated that the snRNAs are associated with hnRNP particles. HnRNP fractions were incubated with normal IgG or anti-U1 RNP IgG, and the resulting immunocomplexes were isolated by binding to a protein A-Sepharose column. HnRNP was found in bound fractions only when anti-U1 RNP IgG was used. By Northern hybridization of RNA extracted from the immunocomplexes with a beta-globin genomic DNA probe, 15S beta-globin mRNA precursors and 10S mature mRNA were detected. These findings suggest the existence of a complex of U1 RNP particles and hnRNP particles containing beta-globin pre-mRNA.

摘要

从弗氏红白血病细胞中分离出不均一核RNA核糖核蛋白（hnRNP）组分，并通过15%-45%蔗糖密度梯度离心进行分离。小核RNA（snRNA）在hnRNP组分中的分布表明，snRNA与hnRNP颗粒相关。将hnRNP组分与正常IgG或抗U1 RNP IgG一起温育，然后通过与蛋白A-琼脂糖柱结合来分离得到的免疫复合物。仅当使用抗U1 RNP IgG时，才在结合组分中发现hnRNP。通过用β-珠蛋白基因组DNA探针与从免疫复合物中提取的RNA进行Northern杂交，检测到了15Sβ-珠蛋白mRNA前体和10S成熟mRNA。这些发现提示存在一种包含β-珠蛋白前体mRNA的U1 RNP颗粒与hnRNP颗粒的复合物。