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Dek42 编码一种 RNA 结合蛋白,该蛋白影响可变剪接前体 mRNA 和玉米籽粒发育。

Dek42 encodes an RNA-binding protein that affects alternative pre-mRNA splicing and maize kernel development.

机构信息

State Key Laboratory of Plant Physiology and Biochemistry, National Maize Improvement Center, Beijing Key Laboratory of Crop Genetic Improvement, Joint International Research Laboratory of Crop Molecular Breeding, College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100193, China.

Shanghai Key Laboratory of Bio-Energy Crops, Plant Science Center, School of Life Sciences, Shanghai University, Shanghai, 200444, China.

出版信息

J Integr Plant Biol. 2019 Jun;61(6):728-748. doi: 10.1111/jipb.12798. Epub 2019 Apr 13.

Abstract

RNA-binding proteins (RBPs) play an important role in post-transcriptional gene regulation. However, the functions of RBPs in plants remain poorly understood. Maize kernel mutant dek42 has small defective kernels and lethal seedlings. Dek42 was cloned by Mutator tag isolation and further confirmed by an independent mutant allele and clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 materials. Dek42 encodes an RRM_RBM48 type RNA-binding protein that localizes to the nucleus. Dek42 is constitutively expressed in various maize tissues. The dek42 mutation caused a significant reduction in the accumulation of DEK42 protein in mutant kernels. RNA-seq analysis showed that the dek42 mutation significantly disturbed the expression of thousands of genes during maize kernel development. Sequence analysis also showed that the dek42 mutation significantly changed alternative splicing in expressed genes, which were especially enriched for the U12-type intron-retained type. Yeast two-hybrid screening identified SF3a1 as a DEK42-interacting protein. DEK42 also interacts with the spliceosome component U1-70K. These results suggested that DEK42 participates in the regulation of pre-messenger RNA splicing through its interaction with other spliceosome components. This study showed the function of a newly identified RBP and provided insights into alternative splicing regulation during maize kernel development.

摘要

RNA 结合蛋白 (RBPs) 在转录后基因调控中发挥着重要作用。然而,RBPs 在植物中的功能仍知之甚少。玉米籽粒突变体 dek42 具有小的缺陷籽粒和致死的幼苗。Dek42 是通过 Mutator 标签分离克隆的,并进一步通过独立的突变等位基因和簇状规则间隔短回文重复 (CRISPR)-CRISPR 相关蛋白 9 材料得到证实。Dek42 编码一种 RRM_RBM48 型 RNA 结合蛋白,定位于细胞核。Dek42 在各种玉米组织中持续表达。Dek42 突变导致突变体籽粒中 DEK42 蛋白积累显著减少。RNA-seq 分析表明,Dek42 突变在玉米籽粒发育过程中显著干扰了数千个基因的表达。序列分析还表明,Dek42 突变显著改变了表达基因的可变剪接,尤其是富含 U12 型内含子保留型。酵母双杂交筛选鉴定出 SF3a1 是 DEK42 的相互作用蛋白。DEK42 还与剪接体成分 U1-70K 相互作用。这些结果表明,DEK42 通过与其他剪接体成分的相互作用参与前信使 RNA 剪接的调节。本研究展示了一种新鉴定的 RBP 的功能,并为玉米籽粒发育过程中的可变剪接调控提供了新的见解。

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