The RNA ligands for mouse proline-rich RNA-binding protein (mouse Prrp) contain two consensus sequences in separate loop structure.
The RNA ligands for mouse proline-rich RNA-binding protein (mouse Prrp) contain two consensus sequences in separate loop structure.
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DOI：
10.1093/nar/gki153
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发表时间：
2005
影响因子：
14.9
通讯作者：
Kurihara Y
中科院分区：
文献类型：
--
作者：
Hori T;Taguchi Y;Uesugi S;Kurihara Y
小鼠富含脯氨酸的RNA结合蛋白（mPrrp）是爪蟾Prrp的一种同源蛋白，与Vg 1 mRNA的3′-非翻译区（3′-UTR）的植物定位元件（VLE）结合，参与Vg 1 mRNA向卵母细胞植物皮质的转运和/或定位。在小鼠睾丸中，mPrrp蛋白在粗线期精母细胞和圆形精子细胞的细胞核中大量表达，并在伸长精子细胞中转移到细胞质中。为了深入了解mPrrp在雄性生殖细胞中的功能，我们进行了体外RNA选择（SELEX），以确定mPrrp的RNA配体序列。分析表明，许多选择的克隆含有两个保守元件，AAAUAG和GU 1 -3AG。对所选RNA的缺失突变体和二级结构分析的RNA结合研究表明，含有保守元件的双环结构是与mPrrp高亲和力结合所必需的。此外，我们还发现爪蟾Prrp的靶基因在3′-UTR中含有完整的AAAUAG和GU 1 -3AG序列，这表明爪蟾Prrp和小鼠Prrp具有相同的结合序列。
Mouse proline-rich RNA-binding protein (mPrrp) is a mouse ortholog of Xenopus Prrp, which binds to a vegetal localization element (VLE) in the 3′-untranslated region (3′-UTR) of Vg1 mRNA and is expected to be involved in the transport and/or localization of Vg1 mRNA to the vegetal cortex of oocytes. In mouse testis, mPrrp protein is abundantly expressed in the nuclei of pachytene spermatocytes and round spermatids, and shifts to the cytoplasm in elongating spermatids. To gain an insight into the function of mPrrp in male germ cells, we performed in vitro RNA selection (SELEX) to determine the RNA ligand sequence of mPrrp. This analysis revealed that many of the selected clones contained both of two conserved elements, AAAUAG and GU1–3AG. RNA-binding study on deletion mutants and secondary structure analyses of the selected RNA revealed that a two-loop structure containing the conserved elements is required for high-affinity binding to mPrrp. Furthermore, we found that the target mRNAs of Xenopus Prrp contain intact AAAUAG and GU1–3AG sequences in the 3′-UTR, suggesting that these binding sequences are shared by Prrps of Xenopus and mouse.
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