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首页> 外文期刊>Biochemistry >The P1/P2 Protein Heterodimers Assemble to the Ribosomal Stalk at the Moment When the Ribosome Is Committed to Translation but Not to the Native 60S Ribosomal Subunit in Saccharomyces cerevisiae
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The P1/P2 Protein Heterodimers Assemble to the Ribosomal Stalk at the Moment When the Ribosome Is Committed to Translation but Not to the Native 60S Ribosomal Subunit in Saccharomyces cerevisiae

机译：P1 / P2蛋白异二聚体组装到核糖体茎的瞬间，当核糖体被翻译但不参与酿酒酵母中天然的60S核糖体亚基时。

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摘要

The four structural acidic ribosomal proteins that dissociate from P1A/P2B and P1B/P2A heterodimers of Saccharomyces cerevisiae were searched in the 60S ribosomal subunit, the 80S monosome, and the polysomal fractions after ribosome profile centrifugation in sucrose gradients in TMN buffer, and after dissociation of monosomes and polysomes to small and large ribosomal subunits in LMS buffer. Analysis by isoelectric focusing, sodium dodecyl sulfate?polyacrylamide gel electrophoresis, and Western blotting of these fractions or the purified acidic protein samples showed eight bands that correspond to the acidic ribosomal proteins in the 60S dissociated subunits of the 80S monosome and polysomes. After samples had been radiolabeled with ~(32)P, four bands were shown to correspond to the phosphorylated form of the acidic ribosomal proteins located in the 80S monosome and the polysomes. Surprisingly, native 60S subunits have no acidic ribosomal proteins. Altogether, these findings indicate that P1/P2 heterodimers bind to P0 when both ribosomal subunits are joined and committed to translation, and they detached from the stalk, just after the small and large ribosomal subunits were separated from the mRNA. Evidence that the phosphorylated and unphosphorylated P1 and P2 acidic ribosomal proteins are part of the functional stalk is also presented.
机译：在60S核糖体亚基，80S单核糖体和多聚体级分中，在核糖体谱中离心蔗糖梯度中的蔗糖梯度离心后，并在解离后，在啤酒酵母的60S核糖体亚基中搜索了四种与酿酒酵母P1A / P2B和P1B / P2A异二聚体解离的酸性结构核糖体蛋白。 LMS缓冲液中单核糖体和多核糖体对大小核糖体亚基的影响。通过等电聚焦分析，十二烷基硫酸钠钠聚丙烯酰胺凝胶电泳以及这些馏分或纯化的酸性蛋白质样品的蛋白质印迹分析，显示了与80S单核糖体和多核糖体的60S解离亚基中的酸性核糖体蛋白质相对应的八个谱带。用〜（32）P放射性标记样品后，显示出四个条带，它们对应于位于80S单核糖体和多核糖体中的酸性核糖体蛋白的磷酸化形式。出乎意料的是，天然的60S亚基没有酸性核糖体蛋白。总而言之，这些发现表明当两个核糖体亚基都与mRNA分离后，当两个核糖体亚基都连接并致力于翻译时，P1 / P2异二聚体就与P0结合，并且它们与茎分离。还提供了磷酸化和非磷酸化的P1和P2酸性核糖体蛋白是功能柄的一部分的证据。

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