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首页> 外文期刊>Cell Research >Developmental expression of Cyclin H and Cdk7 in zebrafish: the essential role of Cyclin H during early embryo development
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Developmental expression of Cyclin H and Cdk7 in zebrafish: the essential role of Cyclin H during early embryo development

机译：Cyclin H和Cdk7在斑马鱼中的发育表达：Cyclin H在早期胚胎发育过程中的重要作用

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摘要

Cyclin-dependent kinase 7 (Cdk7) is the catalytic subunit of the metazoan Cdk-activating kinase (CAK). Activation of Cdk7 requires its association with a regulatory subunit, Cyclin H. Although the Cdk7/Cyclin H complex has been implicated in the regulation of RNA polymerase in several species, the precise function of their orthologs in zebrafish has not been fully elucidated. In this study, we isolated from zebrafish blastula embryos two cDNAs encoding the orthologs of human Cyclin H and Cdk7, and examined the role of Cdkl/Cyclin H in zebrafish embryogenesis. Sequence analysis showed that the zebrafish Cyclin H and Cdk7 cDNAs encode proteins with 65% and 86% identity to the respective human orthologs. RT-PCR and whole-mount in situ hybridization analyses of their expression in unfertilized eggs, embryos and organs of adult fish suggested that Cyclin H and Cdkl messages are maternally loaded. Our data also showed that their transcripts were detected throughout development. Distribution of Cyclin H transcripts was found to be ubiquitous during early stages of development and become restricted to the anterior neural tube, brain, eyes, procreate tissues, liver and heart by 5 days post-fertilization. Expression of a dominant-negative form of Cyclin H delayed the onset of zygotic transcription in the early embryo, resulting in apoptosis at 5 hours post-fertilization and leading to sever defects in tissues normally exhibiting high levels of Cyclin H expression. These results implicate Cyclin H in the regulation of the transcriptional machinery during midblastula transition and suggest that it is an essential gene in early zebrafish larval development.
机译：细胞周期蛋白依赖性激酶7（Cdk7）是后生动物Cdk激活激酶（CAK）的催化亚基。 Cdk7的激活需要将其与调节亚基Cyclin H结合。尽管Cdk7 / Cyclin H复合物与几种物种的RNA聚合酶调控有关，但尚未充分阐明它们在斑马鱼中的直系同源物的精确功能。在这项研究中，我们从斑马鱼囊胚胚胎中分离出两个编码人Cyclin H和Cdk7直系同源基因的cDNA，并研究了Cdkl / Cyclin H在斑马鱼胚胎发生中的作用。序列分析表明，斑马鱼Cyclin H和Cdk7 cDNA编码的蛋白质与各自的人类直系同源物具有65％和86％的同一性。 RT-PCR和成年鱼的未受精卵，胚胎和器官中表达的全量原位杂交分析表明，Cyclin H和Cdkl信息是母源性的。我们的数据还表明，他们的成绩单在整个开发过程中都被检测到。发现Cyclin H转录本的分布在发育的早期阶段普遍存在，并在受精后5天限制在前神经管，大脑，眼睛，生殖组织，肝脏和心脏。 Cyclin H显性阴性形式的表达延迟了早期胚胎中合子转录的开始，导致受精后5小时发生细胞凋亡，并导致通常表现出高水平Cyclin H表达的组织出现严重缺陷。这些结果暗示细胞周期蛋白H在中胚层过渡过程中调控转录机制，并表明它是早期斑马鱼幼虫发育的必需基因。

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