'Edgetic' perturbation of a C. elegans BCL2 ortholog
'Edgetic' perturbation of a C. elegans BCL2 ortholog
复制标题
优雅类 BCL2 同源物的 "边缘 "扰动
DOI：
10.1038/nmeth.1394
复制
发表时间：
2009-11-01
期刊：
影响因子：
48
通讯作者：
Vidal, Marc
中科院分区：
文献类型：
--
作者：
Dreze, Matija;Charloteaux, Benoit;Vidal, Marc
基因和基因产物并不是孤立地发挥作用，而是在高度互连的“相互作用组”网络中发挥作用，该网络被建模为分别代表大分子及其之间相互作用的节点和边的图。我们建议通过有条不紊地使用缺乏单一相互作用的等位基因来研究基因型-表型关联，同时保留所有其他基因，这与旨在完全消除基因产物的遗传方法相反。我们描述了一种基于反向酵母双杂交系统的综合策略，以分离和表征此类边缘特异性或“边缘”等位基因。我们利用 CED-9（一种秀丽隐杆线虫 BCL2 直系同源物）进行了概念验证。使用 ced-9 Edgetic 等位基因，我们发现了细胞凋亡和中心体蛋白之间新的潜在功能联系。这种方法适合更高的通量，特别适用于转基因简单的生物体中的相互作用组网络分析。
Genes and gene products do not function in isolation but within highly interconnected 'interactome' networks, modeled as graphs of nodes and edges representing macromolecules and interactions between them, respectively. We propose to investigate genotype-phenotype associations by methodical use of alleles that lack single interactions, while retaining all others, in contrast to genetic approaches designed to eliminate gene products completely. We describe an integrated strategy based on the reverse yeast two-hybrid system to isolate and characterize such edge-specific, or 'edgetic', alleles. We established a proof of concept with CED-9, a Caenorhabditis elegans BCL2 ortholog. Using ced-9 edgetic alleles, we uncovered a new potential functional link between apoptosis and a centrosomal protein. This approach is amenable to higher throughput and is particularly applicable to interactome network analysis in organisms for which transgenesis is straightforward.