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一种含Brix结构域的核糖体生物发生因子ARPF2及其相互作用蛋白ARRS1的过表达导致[具体生物]的形态变化和寿命延长。

Overexpression of a Brix Domain-Containing Ribosome Biogenesis Factor ARPF2 and its Interactor ARRS1 Causes Morphological Changes and Lifespan Extension in .

作者信息

Maekawa Shugo, Ueda Yoshiaki, Yanagisawa Shuichi

机构信息

Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.

出版信息

Front Plant Sci. 2018 Aug 27;9:1177. doi: 10.3389/fpls.2018.01177. eCollection 2018.

Abstract

The Brix domain is a conserved domain in several proteins involved in ribosome biogenesis in yeast and animals. In the genome, six Brix domain-containing proteins are encoded; however, their molecular functions have not been fully characterized, as yet. Here we report the functional analysis of a Brix domain-containing protein, ARPF2, which is homologous to yeast Rpf2 that plays an essential role in ribosome biogenesis as a component of the 5S ribonucleoprotein particle. By phenotypic characterization of mutants, histochemical GUS staining, and analysis using green fluorescence protein, we show that is an essential and ubiquitously expressed gene encoding a nucleolar protein. Co-immunoprecipitation and split-GFP-based bimolecular fluorescence complementation assays revealed that ARPF2 interacts with a protein named ARRS1, which is homologous to yeast Rrs1 that forms a complex with Rpf2 in yeast. Furthermore, the result of RNA immunoprecipitation assay indicated that ARPF2 interacts with 5S ribosomal RNA (rRNA) or the precursor of 5S rRNA, as well as with the internal transcribed spacer 2 in the precursors of 25S rRNA. Most intriguingly, we found that the overexpression of and leads to characteristic phenotypes, including short stem, abnormal leaf morphology, and long lifespan, in . These results suggest that the function of Brix domain-containing ARPF2 protein in ribosome biogenesis is intimately associated with the growth and development in plants.

摘要

布里克斯结构域是酵母和动物中参与核糖体生物合成的几种蛋白质中的保守结构域。在基因组中,编码了六种含有布里克斯结构域的蛋白质;然而,它们的分子功能尚未完全阐明。在这里,我们报告了一种含有布里克斯结构域的蛋白质ARPF2的功能分析,它与酵母Rpf2同源,Rpf2作为5S核糖核蛋白颗粒的一个组成部分在核糖体生物合成中起重要作用。通过对突变体的表型特征分析、组织化学GUS染色以及使用绿色荧光蛋白进行分析,我们表明ARPF2是一个必需且普遍表达的基因,编码一种核仁蛋白。免疫共沉淀和基于分裂GFP的双分子荧光互补分析表明,ARPF2与一种名为ARRS1的蛋白质相互作用,ARRS1与酵母中的Rrs1同源,Rrs1在酵母中与Rpf2形成复合物。此外,RNA免疫沉淀分析结果表明,ARPF2与5S核糖体RNA(rRNA)或5S rRNA的前体相互作用,也与25S rRNA前体中的内部转录间隔区2相互作用。最有趣的是,我们发现ARPF2和ARRS1的过表达会导致拟南芥出现特征性表型,包括茎短、叶片形态异常和寿命延长。这些结果表明,含有布里克斯结构域的ARPF2蛋白在核糖体生物合成中的功能与植物的生长发育密切相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48a3/6120060/2e8370a6d045/fpls-09-01177-g001.jpg

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