Towards a temperature-sensitive proteome: developing a Drosophila-friendly degron

走向温度敏感的蛋白质组：开发果蝇友好的降解决定子

• 批准号: BB/D019699/1
• 负责人: Cahir O'Kane
• 金额: $ 13.42万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD019699%2F1
• 关键词: Towards; temperature; sensitive; proteome; developing

Here we propose to develop a new technology that enables biologists to turn activity of genes on and off simply by changing the temperature, by targeting their products for degradation only at higher culture temperatures. This technology is based on well-established biological mechanisms employed by the cell to achieve regulated removal of unwanted proteins. This regulated degradation is mediated by the attachment of degradation signals to the target. Some proteins (degrons), identified in yeast, can induce attachment of these signals to themselves when their structure is disrupted at higher temperatures, thus inducing degradation of themselves and any fusion proteins that they are part of. We and others have successfully shown proof-of-principle that the system can be used in a more complex model organism, Drosophila (fruitfly). However, the temperatures at which degradation of the target is induced in yeast are not well-tolerated by Drosophila, or by other 'cold-blooded' organisms. Therefore, we propose to isolate novel variants of the yeast degron signal, that work better in the normal temperature ranges of Drosophila and of other model organisms widely used by biologists, and to test these in cultured Drosophila cells and in genetically manipulated flies.

在这里，我们建议开发一种新技术，使生物学家能够通过改变温度来打开和关闭基因的活性，目标是使他们的产物仅在较高的培养温度下降解。该技术基于细胞采用的完善的生物机制，以实现调节去除不需要的蛋白质。这种受调节的降解是通过将降解信号附着到靶标来介导的。在酵母中发现的一些蛋白质（降解决定子），当其结构在较高温度下被破坏时，可以诱导这些信号附着到自身上，从而诱导其自身以及它们所属的任何融合蛋白的降解。我们和其他人已经成功地证明了该系统可以用于更复杂的模型生物——果蝇（果蝇）。然而，果蝇或其他“冷血”生物体不能很好地耐受酵母中诱导靶标降解的温度。因此，我们建议分离酵母降解决定子信号的新变体，这些变体在果蝇和生物学家广泛使用的其他模式生物的正常温度范围内工作得更好，并在培养的果蝇细胞和基因操纵的果蝇中测试这些变体。

项目成果

A phagocytic route for uptake of double-stranded RNA in RNAi.

RNAi 中摄取双链 RNA 的吞噬途径。

• DOI: 10.1371/journal.pone.0019087
• 发表时间: 2011-04-29
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Rocha JJ;Korolchuk VI;Robinson IM;O'Kane CJ
• 通讯作者: O'Kane CJ