McGrath K E, Smothers J F, Dadd C A, Madireddi M T, Gorovsky M A, Allis C D
Department of Biology, University of Rochester, New York 14627, USA.
Mol Biol Cell. 1997 Jan;8(1):97-108. doi: 10.1091/mbc.8.1.97.
An abundant 52-kDa phosphoprotein was identified and characterized from macronuclei of the ciliated protozoan Tetrahymena thermophila. Immunoblot analyses combined with light and electron microscopic immunocytochemistry demonstrate that this polypeptide, termed Nopp52, is enriched in the nucleoli of transcriptionally active macronuclei and missing altogether from transcriptionally inert micronuclei. The cDNA sequence encoding Nopp52 predicts a polypeptide whose amino-terminal half consists of multiple acidic/serine-rich regions alternating with basic/proline-rich regions. Multiple serines located in these acidic stretches lie within casein kinase II consensus motifs, and Nopp52 is an excellent substrate for casein kinase II in vitro. The carboxyl-terminal half of Nopp52 contains two RNA recognition motifs and an extreme carboxyl-terminal domain rich in glycine, arginine, and phenylalanine, motifs common in many RNA processing proteins. A similar combination and order of motifs is found in vertebrate nucleolin and yeast NSR1, suggesting that Nopp52 is a member of a family of related nucleolar proteins. NSR1 and nucleolin have been implicated in transcriptional regulation of rDNA and rRNA processing. Consistent with a role in ribosomal gene metabolism, rDNA and Nopp52 colocalize in situ, as well as by cross-linking and immunoprecipitation experiments, demonstrating an association between Nopp52 and rDNA in vivo.
从嗜热栖热四膜虫的大核中鉴定并表征了一种丰富的52-kDa磷蛋白。免疫印迹分析结合光镜和电镜免疫细胞化学表明,这种被称为Nopp52的多肽在转录活跃的大核核仁中富集,而在转录惰性的小核中则完全缺失。编码Nopp52的cDNA序列预测的多肽,其氨基末端一半由多个富含酸性/丝氨酸的区域与富含碱性/脯氨酸的区域交替组成。位于这些酸性区域的多个丝氨酸位于酪蛋白激酶II共有基序内,并且Nopp52在体外是酪蛋白激酶II的优良底物。Nopp52的羧基末端一半包含两个RNA识别基序和一个富含甘氨酸、精氨酸和苯丙氨酸的极端羧基末端结构域,这些基序在许多RNA加工蛋白中常见。在脊椎动物核仁素和酵母NSR1中发现了类似的基序组合和顺序,表明Nopp52是相关核仁蛋白家族的成员。NSR1和核仁素与rDNA的转录调控和rRNA加工有关。与在核糖体基因代谢中的作用一致,rDNA和Nopp52在原位共定位,以及通过交联和免疫沉淀实验,证明了Nopp52与体内rDNA之间的关联。