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The analysis of dharma/bozozok gene function in vertebrates body axis formation.

dharma/bozozok 基因在脊椎动物体轴形成中的功能分析。

基本信息

批准号：
13670013
负责人：
SHIMIZU Takashi
金额：
$ 2.3万
依托单位：
RIKEN (2002)Osaka University (2001)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670013/
关键词：
Organizer Wnt dhama bozozok Homeobox Transcriptional repressor vox vent ved Axis formation 脊椎動物ゼブラフィッシュ boz vega

项目摘要

Specification of dorso-ventral body axis requires the maternally-derived Wnt signal activation. The maternally-derived Wnt signal activates genes that are involved in formation of the Nieuwkoop center and subsequently induction of the dorsal organizer. The homeobox gene dharma/bozozok (dha/boz) is a direct target gene of the maternal Wnt signal and plays an important role in the formation and/or function of the zebra fish dorsal organizer. Dha/Boz contains an amino acid motif similar to the repressor domain of Engrailed (Ehl motif). We found that Dha/Boz interacted with a co-repressor Groucho through the Eh1 motif. The repressor activity of Dha/Boz was required for expression of organizer genes and dha/boz-dependent dorsalization of the embryos. By ordered differential display method, we isolated a novel ventrally-expressed homeobox gene ved, which displays sequence similarities with vox and vent, ved is a target of the transcriptional repressors Dha/Boz. The analysis of transcriptional activator and repressor domain fusion proteins suggested that Ved acts as a transcriptional repressor. By antisense morpholino-mediated loss of function experiments, we showed that ved, vox and vent were redundant repressors of the dorsal organizer formation. These results suggested that dha/boz promoted the dorsal fate by suppressing the expression of ved, vox, and vent which act to restrict the expression of the dorsal organizer genes in appropriate region.

背侧-腹侧轴的规范需要母体来源的Wnt信号激活。母体来源的Wnt信号激活了与Nieuwkoop中心的形成以及随后的背部组织者的诱导有关的基因。同源框基因dharma/bozozok(dha/boz)是母体Wnt信号的直接靶基因，在斑马鱼背部组织者的形成和/或功能中发挥重要作用。DHA/BOZ含有一个与Engraded(Ehl基序)抑制域相似的氨基酸基序。我们发现DHA/BOZ通过EH1基序与共抑制因子Groucho相互作用。组织者基因的表达和依赖于DHA/BOZ的胚胎背化需要DHA/BOZ的阻遏活性。通过有序差异显示的方法，我们克隆了一个新的腹部表达的同源框基因Ved，该基因与VOX和Vent具有序列相似性，Ved是转录抑制因子DHA/Boz的靶标。对转录激活域和抑制域融合蛋白的分析表明，Ved是一种转录抑制域。通过反义吗啉介导的功能丧失实验，我们证明Ved、VOX和Vent是背部组织者形成的多余阻滞物。这些结果表明，DHA/BOZ通过抑制Ved、VOX和Vent的表达来促进背部命运，这些表达在适当的区域限制了背部组织者基因的表达。