DEAD-box protein Ddx46 is required for the development of the digestive organs and brain in zebrafish.
DEAD-box protein Ddx46 is required for the development of the digestive organs and brain in zebrafish.
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DOI：
10.1371/journal.pone.0033675
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kikuchi Y
中科院分区：
文献类型：
--
作者：
Hozumi S;Hirabayashi R;Yoshizawa A;Ogata M;Ishitani T;Tsutsumi M;Kuroiwa A;Itoh M;Kikuchi Y
空间和时间控制的基因表达，包括转录，几个mRNA加工步骤，成熟mRNA的出口到细胞质，是必不可少的发育过程。众所周知，DExD/H-box蛋白家族的RNA解旋酶参与这些基因表达过程，包括转录、前mRNA剪接和rRNA生物发生。虽然一种DExD/H-box蛋白Prp 5（脊椎动物Ddx 46的同源物）已被证明在酵母中的前体mRNA剪接中起重要作用，但Ddx 46在后生动物中的体内功能仍有待充分阐明。在这项研究中，我们分离出斑马鱼morendo（莫尔），这是一种在消化器官和大脑中显示发育缺陷的突变体，并发现它编码Ddx 46。Ddx 46转录本是由母体提供的，随着斑马鱼幼虫的发育，其普遍存在的表达逐渐局限于这些器官。整装原位杂交的结果表明，在这些器官中的各种分子标记的表达显着减少Ddx 46突变体。此外，剪接状态分析与RT-PCR显示未剪接形式的mRNA在消化器官和脑组织的Ddx 46突变体，表明Ddx 46可能需要在斑马鱼发育过程中的前mRNA剪接。因此，我们的研究结果表明，斑马鱼Ddx 46可能通过控制前体mRNA剪接来促进消化器官和大脑的发育。
Spatially and temporally controlled gene expression, including transcription, several mRNA processing steps, and the export of mature mRNA to the cytoplasm, is essential for developmental processes. It is well known that RNA helicases of the DExD/H-box protein family are involved in these gene expression processes, including transcription, pre-mRNA splicing, and rRNA biogenesis. Although one DExD/H-box protein, Prp5, a homologue of vertebrate Ddx46, has been shown to play important roles in pre-mRNA splicing in yeast, the in vivo function of Ddx46 remains to be fully elucidated in metazoans. In this study, we isolated zebrafish morendo (mor), a mutant that shows developmental defects in the digestive organs and brain, and found that it encodes Ddx46. The Ddx46 transcript is maternally supplied, and as development proceeds in zebrafish larvae, its ubiquitous expression gradually becomes restricted to those organs. The results of whole-mount in situ hybridization showed that the expression of various molecular markers in these organs is considerably reduced in the Ddx46 mutant. Furthermore, splicing status analysis with RT-PCR revealed unspliced forms of mRNAs in the digestive organ and brain tissues of the Ddx46 mutant, suggesting that Ddx46 may be required for pre-mRNA splicing during zebrafish development. Therefore, our results suggest a model in which zebrafish Ddx46 is required for the development of the digestive organs and brain, possibly through the control of pre-mRNA splicing.
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