大鼠肝脏信使核糖核蛋白结构的研究。2非poly(A) RNase抗性片段和mRNP的电镜外观。

T Tomcsányi, L Komáromy, A Tigyi
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引用次数: 0

摘要

用5(微克/毫升或0.1微克/毫升胰核糖核酸酶(EC 3.1.27.5)处理部分片段化的[3H]-椰浆酸或[3H]-腺嘌呤标记的12-21S大鼠肝脏信使核糖核蛋白(mRNP),用蔗糖梯度离心分离抗性片段。获得了两种类型的碎片。核糖核酸酶以5微克/毫升的浓度消化mRNP,产生9S多聚(a)- mRNP蛋白颗粒,其特征沉淀,RNA片段的电泳迁移率和蛋白质组成证明了这一点。相比之下,0.1微克/毫升浓度的核糖核酸酶产生2-5S嘧啶标记的片段,携带由大约16-50个残基组成的多核苷酸序列。在这些序列上发现了两个分子量分别为38 000 (P38)和44 000 (P44)的大鼠肝脏mRNP多肽。这些数据表明,在mRNP中,除了poly(A)外,RNA序列还与蛋白质相互作用。电镜研究表明,EDTA将mRNP从多体中解放出来,改变了其表面性质,因为EDTA主要以球状颗粒的形式释放mRNP。然而,一小部分mRNP仍保持原纤维样形式。在原纤维中可以看到几个斑点,可能是附着在mRNA上的蛋白质。根据现有的数据,提出了一个“平均”mRNP分子的结构模型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Studies on the structure of rat liver messenger ribonucleoprotein. II. Non-poly(A) RNase resistant fragments and electron microscopic appearance of mRNP.

Partially fragmented 12-21S rat liver messenger ribonucleoprotein (mRNP), labelled either with [3H]-orotic acid or [3H]-adenine was treated with 5 (micrograms/ml or 0.1 microgram/ml pancreatic ribonuclease (EC 3.1.27.5) and the resistant fragments were separated by sucrose gradient centrifugation. Two types of fragments were obtained. Digestion of mRNP with ribonuclease at a concentration of 5 micrograms/ml resulted in 9S poly(A)-protein particles of mRNP as evidenced by their characteristic sedimentation, electrophoretic mobility of the RNA moiety and protein composition. In contrast, ribonuclease at a concentration of 0.1 microgram/ml produced 2-5S pyrimidine labelled fragments carrying polynucleotide sequences consisting of approximately 16-50 residues. Two polypeptides of rat liver mRNP with molecular weights of 38 000 (P38) and 44 000 (P44) were found to be attached to these sequences. The data demonstrate that RNA sequences other than poly(A) interact with protein in mRNP. Electron microscopic studies revealed that the liberation of mRNP from the polysomes by EDTA changes its surface properties since EDTA releases the mRNP mainly in the form of globular particles. However, a small proportion of the mRNP remains in fibril-like form. Among the fibrils several blobs could be seen which were probably the proteins attached to the mRNA. From the available data a model for the structure of an "average" mRNP molecule is proposed.

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