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APUM23对18S rRNA特异性识别的结构基础

Structural basis for the specific recognition of 18S rRNA by APUM23.

作者信息

Bao Hongyu, Wang Na, Wang Chongyuan, Jiang Yiyang, Liu Jiuyang, Xu Li, Wu Jihui, Shi Yunyu

机构信息

Hefei National Laboratory for Physical Science at Microscale, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China.

CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

出版信息

Nucleic Acids Res. 2017 Nov 16;45(20):12005-12014. doi: 10.1093/nar/gkx872.

DOI:10.1093/nar/gkx872
PMID:29036323
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5714250/
Abstract

PUF (Pumilio/fem-3 mRNA binding factor) proteins, a conserved family of RNA-binding proteins, recognize specific single-strand RNA targets in a specific modular way. Although plants have a greater number of PUF protein members than do animal and fungal systems, they have been the subject of fewer structural and functional investigations. The aim of this study was to elucidate the involvement of APUM23, a nucleolar PUF protein in the plant Arabidopsis, in pre-rRNA processing. APUM23 is distinct from classical PUF family proteins, which are located in the cytoplasm and bind to 3'UTRs of mRNA to modulate mRNA expression and localization. We found that the complete RNA target sequence of APUM23 comprises 11 nt in 18S rRNA at positions 1141-1151. The complex structure shows that APUM23 has 10 PUF repeats; it assembles into a C-shape, with an insertion located within the inner concave surface. We found several different RNA recognition features. A notable structural feature of APUM23 is an insertion in the third PUF repeat that participates in nucleotide recognition and maintains the correct conformation of the target RNA. Our findings elucidate the mechanism for APUM23's-specific recognition of 18S rRNA.

摘要

PUF（Pumilio/fem-3 mRNA结合因子）蛋白是一类保守的RNA结合蛋白家族，它们以特定的模块化方式识别特定的单链RNA靶标。尽管植物中的PUF蛋白成员比动物和真菌系统中的更多，但对它们进行的结构和功能研究却较少。本研究的目的是阐明拟南芥中的一种核仁PUF蛋白APUM23在rRNA前体加工中的作用。APUM23与经典的PUF家族蛋白不同，经典PUF家族蛋白位于细胞质中，与mRNA的3'非翻译区结合以调节mRNA的表达和定位。我们发现APUM23的完整RNA靶标序列由18S rRNA中第1141 - 1151位的11个核苷酸组成。复杂结构显示APUM23有10个PUF重复序列；它组装成C形，在内部凹面有一个插入片段。我们发现了几种不同的RNA识别特征。APUM23一个显著的结构特征是在第三个PUF重复序列中有一个插入片段，该片段参与核苷酸识别并维持靶标RNA的正确构象。我们的研究结果阐明了APUM23特异性识别18S rRNA的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/ccee07120eb3/gkx872fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/074f1412ca60/gkx872fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/f1b4125d4999/gkx872fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/4f668fed86ba/gkx872fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/b35c8e893253/gkx872fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/68fc32ed1a5d/gkx872fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/ccee07120eb3/gkx872fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/074f1412ca60/gkx872fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/f1b4125d4999/gkx872fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/4f668fed86ba/gkx872fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/b35c8e893253/gkx872fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/68fc32ed1a5d/gkx872fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d44/5714250/ccee07120eb3/gkx872fig6.jpg

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Nat Commun. 2016 Oct 11;7:13085. doi: 10.1038/ncomms13085.
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Programmable RNA-binding protein composed of repeats of a single modular unit.由单个模块化单元重复序列组成的可编程RNA结合蛋白。
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Distinct RNA-binding modules in a single PUF protein cooperate to determine RNA specificity.单个 PUF 蛋白中的独特 RNA 结合模块协同合作以确定 RNA 特异性。
Nucleic Acids Res. 2019 Sep 19;47(16):8770-8784. doi: 10.1093/nar/gkz583.
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