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首页> 外文期刊>Journal of Molecular Biology >Ribosomal RNA maturation in Schizosaccharomyces pombe is dependent on a large ribonucleoprotein complex of the internal transcribed spacer 1.
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Ribosomal RNA maturation in Schizosaccharomyces pombe is dependent on a large ribonucleoprotein complex of the internal transcribed spacer 1.

机译：粟酒裂殖酵母中核糖体RNA的成熟取决于内部转录间隔区1的大型核糖核蛋白复合物。

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摘要

The interdependency of steps in the processing of pre-rRNA in Schizosaccharomyces pombe suggests that RNA processing, at least in part, acts as a quality control mechanism which helps assure that only functional RNA is incorporated into mature ribosomes. To determine further the role of the transcribed spacer regions in rRNA processing and to detect interactions which underlie the interdependencies, the ITS1 sequence was examined for its ability to form ribonucleoprotein complexes with cellular proteins. When incubated with protein extract, the spacer formed a specific large RNP. This complex was stable to fractionation by agarose or polyacrylamide gel electrophoresis. Modification exclusion analyses indicated that the proteins interact with a helical domain which is conserved in the internal transcribed spacers. Mutagenic analyses confirmed an interaction with this sequence and indicated that this domain is critical to the efficient maturation of the precursor RNA. The protein constituents, purified by affinity chromatography using the ITS1 sequence, retained an ability to form stable RNP. Protein analyses of gel purified complex, prepared with affinity-purified proteins, indicated at least 20 protein components ranging in size from 20-200 kDa. Peptide mapping by Maldi-Toff mass spectroscopy identified eight hypothetical RNA binding proteins which included four different RNA-binding motifs. Another protein was putatively identified as a pseudouridylate synthase. Additional RNA constituents were not detected. The significance of this complex with respect to rRNA maturation and interdependence in rRNA processing is discussed. Copyright 2000 Academic Press.
机译：粟酒裂殖酵母中前rRNA加工步骤之间的相互依赖性表明，RNA加工至少部分充当质量控制机制，有助于确保仅将功能性RNA掺入成熟核糖体中。为了进一步确定转录间隔区在rRNA加工中的作用并检测相互依赖的相互作用，检查了ITS1序列与细胞蛋白形成核糖核蛋白复合物的能力。与蛋白质提取物一起孵育时，间隔基形成了特定的大RNP。该复合物对于通过琼脂糖或聚丙烯酰胺凝胶电泳分级分离是稳定的。修饰排除分析表明，蛋白质与内部转录的间隔区中保守的螺旋结构域相互作用。诱变分析证实了与该序列的相互作用，并表明该结构域对前体RNA的有效成熟至关重要。通过使用ITS1序列的亲和层析纯化的蛋白质成分，保留了形成稳定RNP的能力。用亲和纯化的蛋白质制备的凝胶纯化的复合物的蛋白质分析表明，至少20种蛋白质成分的大小在20-200 kDa之间。通过Maldi-Toff质谱进行的肽图分析鉴定了八个假设的RNA结合蛋白，其中包括四个不同的RNA结合基序。推定另一种蛋白质为假尿苷酸合酶。未检测到其他RNA成分。讨论了该复合物在rRNA加工中对于rRNA成熟和相互依赖性的重要性。版权所有2000学术出版社。

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