Endo Y, Tominaga H, Natori Y
Biochim Biophys Acta. 1975 Mar 21;383(3):305-15. doi: 10.1016/0005-2787(75)90059-3.
The administration of ethionine to female rats causes breakdown of hepatic polysomes. The state of mRNA and monomeric ribosomes after the polysome dissociation was studied. The mRNA was selectively labeled with [14C] orotate after a low dose of actinomycin D. Sucrose density gradient centrifugation of Triton X-100-treated cytoplasm revealed an accumulation of heterodisperse radioactive material with very large S values. This material was converted to smaller S values with deoxycholate treatment and was extremely sensitive to mild ribonuclease treatment. Since this material was banded at around 1.43 g/cm3 in CsCl gradient centrifugation and contained RNA with a distribution of S values characteristic of polysomal mRNA, this material was identified as mRNA-containing ribonucleoprotein particles. The monomeric ribosomes were shown to be dissociated into subunits in the presence of 0.5 M KCl, indicating that these lacked nascent polypeptide chains. When the animals were recovered from the ethionine treatment by subsequent administration of adenine and methionine, the heterodisperse ribonucleoprotein particles and monomeric ribosomes appeared to be utilized for the reformation of polysomes.
给雌性大鼠施用乙硫氨酸会导致肝多核糖体的解体。研究了多核糖体解离后mRNA和单体核糖体的状态。在低剂量放线菌素D处理后,用[14C]乳清酸对mRNA进行选择性标记。对经 Triton X - 100处理的细胞质进行蔗糖密度梯度离心,发现积累了具有非常大S值的多分散放射性物质。用脱氧胆酸盐处理后,这种物质转化为较小的S值,并且对温和的核糖核酸酶处理极为敏感。由于这种物质在CsCl梯度离心中在约1.43 g/cm3处形成条带,并且含有具有多核糖体mRNA特征性S值分布的RNA,因此这种物质被鉴定为含mRNA的核糖核蛋白颗粒。结果表明,在0.5 M KCl存在下,单体核糖体解离成亚基,这表明这些亚基缺乏新生多肽链。当通过随后施用腺嘌呤和蛋氨酸使动物从乙硫氨酸处理中恢复时,多分散核糖核蛋白颗粒和单体核糖体似乎被用于多核糖体的重新形成。