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首页> 美国卫生研究院文献>Molecular and Cellular Biology >Bop1 Is a Mouse WD40 Repeat Nucleolar Protein Involved in 28S and 5.8S rRNA Processing and 60S Ribosome Biogenesis
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Bop1 Is a Mouse WD40 Repeat Nucleolar Protein Involved in 28S and 5.8S rRNA Processing and 60S Ribosome Biogenesis

机译：Bop1是涉及28S和5.8S rRNA加工和60S核糖体生物发生的小鼠WD40重复核仁蛋白。

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We have identified and characterized a novel mouse protein, Bop1, which contains WD40 repeats and is highly conserved through evolution. bop1 is ubiquitously expressed in all mouse tissues examined and is upregulated during mid-G1 in serum-stimulated fibroblasts. Immunofluorescence analysis shows that Bop1 is localized predominantly to the nucleolus. In sucrose density gradients, Bop1 from nuclear extracts cosediments with the 50S-80S ribonucleoprotein particles that contain the 32S rRNA precursor. RNase A treatment disrupts these particles and releases Bop1 into a low-molecular-weight fraction. A mutant form of Bop1, Bop1Δ, which lacks 231 amino acids in the N- terminus, is colocalized with wild-type Bop1 in the nucleolus and in ribonucleoprotein complexes. Expression of Bop1Δ leads to cell growth arrest in the G1 phase and results in a specific inhibition of the synthesis of the 28S and 5.8S rRNAs without affecting 18S rRNA formation. Pulse-chase analyses show that Bop1Δ expression results in a partial inhibition in the conversion of the 36S to the 32S pre-rRNA and a complete inhibition of the processing of the 32S pre-rRNA to form the mature 28S and 5.8S rRNAs. Concomitant with these defects in rRNA processing, expression of Bop1Δ in mouse cells leads to a deficit in the cytosolic 60S ribosomal subunits. These studies thus identify Bop1 as a novel, nonribosomal mammalian protein that plays a key role in the formation of the mature 28S and 5.8S rRNAs and in the biogenesis of the 60S ribosomal subunit.
机译：我们已经鉴定并鉴定了一种新型小鼠蛋白Bop1，它含有WD40重复序列，并且在进化过程中高度保守。 bop1在所有检查的小鼠组织中普遍表达，并且在血清刺激的成纤维细胞的G1中期被上调。免疫荧光分析表明Bop1主要位于核仁。在蔗糖密度梯度中，核提取物中的Bop1与含有32S rRNA前体的50S-80S核糖核蛋白颗粒共沉淀。 RNase A处理会破坏这些颗粒并将Bop1释放为低分子量部分。在N端缺少231个氨基酸的Bop1突变体Bop1Δ与野生型Bop1在核仁和核糖核蛋白复合物中共定位。 Bop1Δ的表达导致细胞生长停滞在G1期，并导致28S和5.8S rRNA合成的特异性抑制，而不会影响18S rRNA的形成。脉冲追踪分析表明，Bop1Δ的表达导致36S向32S pre-rRNA的转化受到部分抑制，并完全抑制32S pre-rRNA形成成熟的28S和5.8S rRNA的过程。伴随rRNA加工中的这些缺陷，小鼠细胞中Bop1Δ的表达导致胞质60S核糖体亚基的缺乏。因此，这些研究将Bop1鉴定为一种新型的非核糖体哺乳动物蛋白，它在成熟的28S和5.8S rRNA的形成以及60S核糖体亚基的生物发生中起关键作用。

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