Functional analysis of a novel small peptide gene in Drosophila pattern formation

果蝇模式形成中新型小肽基因的功能分析

• 批准号: 18570200
• 负责人: KAGEYAMA Yuji
• 金额: $ 2.63万
• 依托单位: National Institutes of Natural Sciences Okazaki Research Facilities (2007)Nara Institute of Science and Technology (2006)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18570200/
• 关键词: Drosophila; organogenesis; transcriptional regulation; cytoskeleton; cell architecture; denticle; small peptide; actin; キイロショウジョウバエ; 上皮細胞

Denticle is a special cuticle structure of larval epithelial cells in Drosophila, which is prepatterned by protrusion of embryonic epidermal cells. In previous studies, we have shown that the polished rice(pri) gene plays an important role in denticle formation. To elucidate molecular basis of pattern formation in Drosophila, we characterized the pri gene by a series of genetic experiments.pri transcript does not contains any ORFs longer than 49 aa, but five of ten short ORFs are evolutionarily conserved. Reporter assays using series of pri transgenes showed that the first four ORFs(11, 11, 11 and 32 aa), which share a common septamer motif LDPTGQ/TY, were independently translated from the full-length pri cDNA transgenes. When each of any four ORFs was introduced in pri mutants, the denticle phenotype was completely rescued. On the other hands, a control transgene containing frame-shift mutation did not show any effects on the phenotype. These results suggest that pri encode multiple small peptides polycistronically, which share redundant function during denticle formation.By time-lapse imaging analysis, we found that pri mutant shows evident defects of actin dynamics during denticle formation. In control embryos, intracellular localization of actin fibers prefigured cell protrusions in denticle cells, while pri mutants did not show any sign of the localized signal. We also found that pri is required for expression of several actin regulators controlled by transcriptional factor Shavenbaby(SVB), although expression of SVB itself is not affected by pri mutation, suggesting that pri is involved in transcriptional regulation by SVB protein.Taken together, we conclude that pri encodes multiple small peptides that regulate denticle formation via intercellular interaction. Taking account of the fact that eukaryotic genomes contain a number of short ORF genes, our results suggest that potent importance of small peptide genes in developmental events.

细齿是果蝇幼虫上皮细胞的一种特殊角质层结构，是由胚胎表皮细胞突出预先形成的。在之前的研究中，我们已经证明精米（pri）基因在小齿形成中发挥着重要作用。为了阐明果蝇模式形成的分子基础，我们通过一系列遗传实验对 pri 基因进行了表征。pri 转录本不包含任何长于 49 个氨基酸的 ORF，但十个短 ORF 中有五个在进化上是保守的。使用一系列 pri 转基因的报告基因检测表明，前四个 ORF（11、11、11 和 32 个氨基酸）具有共同的七聚体基序 LDPTGQ/TY，是从全长 pri cDNA 转基因独立翻译的。当任意四个 ORF 中的每一个被引入 pri 突变体时，小齿表型都被完全挽救。另一方面，含有移码突变的对照转基因没有显示出对表型的任何影响。这些结果表明pri以多顺反子的方式编码多个小肽，这些小肽在小齿形成过程中共享冗余功能。通过延时成像分析，我们发现pri突变体在小齿形成过程中表现出明显的肌动蛋白动力学缺陷。在对照胚胎中，肌动蛋白纤维的细胞内定位预示了小齿细胞中的细胞突起，而 pri 突变体没有显示出任何定位信号的迹象。我们还发现pri是转录因子Shavenbaby（SVB）控制的几种肌动蛋白调节因子的表达所必需的，尽管SVB本身的表达不受pri突变的影响，这表明pri参与了SVB蛋白的转录调节。综上所述，我们得出结论，pri编码多个小肽，通过细胞间相互作用调节齿状突的形成。考虑到真核基因组含有许多短 ORF 基因这一事实，我们的结果表明小肽基因在发育事件中具有潜在的重要性。

项目成果

Drosophila blimp-1 is a transient transcriptional repressor that controls timing of the ecdysone-induced developmental pathway

• DOI: 10.1128/mcb.01304-07
• 发表时间: 2007-12-01
• 期刊: MOLECULAR AND CELLULAR BIOLOGY
• 影响因子: 5.3
• 作者: Agawa, Yasuo;Sarhan, Moustafa;Ueda, Hitoshi
• 通讯作者: Ueda, Hitoshi

The role of a short peptide gene, polished rice, during imaginal development.

短肽基因（精米）在图像发育过程中的作用。

• 发表时间: 2007
• 作者: Yoshiko Hashimoto;Kondo;T. and Kageyama;Y.
• 通讯作者: Y.

Polycistronically-encoded small peptides regulate actin-based morphogenesis in Drosophila.

多顺反子编码的小肽调节果蝇中基于肌动蛋白的形态发生。

• 发表时间: 2007
• 作者: Takefumi Kondo;Kato;K.;Hashimoto;Y.;Hayashi;S. and Kageyama;Y.
• 通讯作者: Y.

Short peptide gene,polished rice,regulate Drosophila imaginal development via Cell-Cell intelaction

短肽基因、精米、细胞间相互作用调控果蝇成像发育

• 发表时间: 2007
• 作者: Yoshiko Hashimoto;Kondo;T. and Kageyama;Y.
• 通讯作者: Y.

Polycistronically-encoded Small Peptides Regulate Actin-based Epithelial Morphol ogy In Drosophila

多顺反子编码的小肽调节果蝇中基于肌动蛋白的上皮形态

• 发表时间: 2007
• 作者: Takeshi Fukuyama;Tatsuru Kikuchi and Nobuchika Okada;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Tatsuru Kikuchi;Takefumi Kondo;Takefumi Kondo;近藤武史;近藤武史;Takefumi Kondo
• 通讯作者: Takefumi Kondo