Systematic screening of a Drosophila ORF library in vivo uncovers Wnt/Wg pathway components.
Systematic screening of a Drosophila ORF library in vivo uncovers Wnt/Wg pathway components.
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DOI：
10.1016/j.devcel.2013.02.019
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发表时间：
2013-04
影响因子：
11.8
通讯作者：
C. Schertel;Dashun Huang;M. Björklund;J. Bischof;Dingzi Yin;Rongxia Li;Yi Wu;Rong Zeng;Jiarui Wu;J. Taipale;Haiyun Song;K. Basler
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文献类型：
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作者：
C. Schertel;Dashun Huang;M. Björklund;J. Bischof;Dingzi Yin;Rongxia Li;Yi Wu;Rong Zeng;Jiarui Wu;J. Taipale;Haiyun Song;K. Basler
我们在果蝇中创建了一个包含 655 个生长调节基因的定点 UAS-ORF 文库。该文库代表了大量调节细胞周期、细胞大小和增殖的基因，将成为研究体内生长调节的宝贵资源。通过使用基因的错误表达，我们可以防止遗传冗余引起的问题，并可以发现新的基因功能。为了验证该库的实用性，我们筛选了 Wingless (Wg) 通路组件。我们结合实验和生物信息学方法来预测候选者，并确定了三种丝氨酸/苏氨酸激酶作为 Wg 信号传导的调节剂。我们证明其中之一 Nek2 通过直接磷酸化 Dishevelled 来优化途径反应。此外，我们描述了大约 5% 的果蝇基因的功能关系，并鉴定了大量在错误表达时调节细胞大小、增殖和最终器官大小的基因。
We created a site-directedUAS-ORFlibrary of 655 growth-regulating genes inDrosophila. This library represents a large collection of genes regulating cell cycle, cell size, and proliferation and will be a valuable resource for studying growth regulation in vivo. By using misexpression of genes, we prevent problems arising from genetic redundancy and can uncover novel gene functions. To validate the usefulness of this library, we screened for Wingless (Wg) pathway components. We used a combination of experimental and bioinformatic approaches to predict candidates and identified three serine/threonine kinases as regulators of Wg signaling. We show that one of these, Nek2, optimizes pathway response by direct phosphorylation of Dishevelled. In addition, we describe functional relations for roughly 5% of allDrosophilagenes and identify a large number of genes that regulate cell size, proliferation, and final organ size upon misexpression.