Department of Molecular Genetics, The Donnelly Centre, University of Toronto, Toronto, Ontario M5S3E1, Canada.
Genome Res. 2012 Apr;22(4):791-801. doi: 10.1101/gr.129213.111. Epub 2012 Jan 26.
A combinatorial genetic perturbation strategy was applied to interrogate the yeast kinome on a genome-wide scale. We assessed the global effects of gene overexpression or gene deletion to map an integrated genetic interaction network of synthetic dosage lethal (SDL) and loss-of-function genetic interactions (GIs) for 92 kinases, producing a meta-network of 8700 GIs enriched for pathways known to be regulated by cognate kinases. Kinases most sensitive to dosage perturbations had constitutive cell cycle or cell polarity functions under standard growth conditions. Condition-specific screens confirmed that the spectrum of kinase dosage interactions can be expanded substantially in activating conditions. An integrated network composed of systematic SDL, negative and positive loss-of-function GIs, and literature-curated kinase-substrate interactions revealed kinase-dependent regulatory motifs predictive of novel gene-specific phenotypes. Our study provides a valuable resource to unravel novel functional relationships and pathways regulated by kinases and outlines a general strategy for deciphering mutant phenotypes from large-scale GI networks.
采用组合遗传干扰策略在全基因组范围内研究酵母激酶组。我们评估了基因过表达或基因缺失的全局效应，以绘制 92 种激酶的合成剂量致死（SDL）和功能丧失遗传相互作用（GI）的综合遗传相互作用网络，生成了一个包含 8700 个 GI 的元网络，这些 GI 富集了已知受同源激酶调节的途径。在标准生长条件下，对剂量扰动最敏感的激酶具有组成性细胞周期或细胞极性功能。条件特异性筛选证实，在激活条件下，激酶剂量相互作用的范围可以大大扩展。由系统 SDL、负和正功能丧失 GI 以及文献整理的激酶-底物相互作用组成的综合网络揭示了激酶依赖性调节基序，可预测新的基因特异性表型。我们的研究提供了一个有价值的资源，可以揭示激酶调节的新的功能关系和途径，并概述了从大规模 GI 网络中解码突变表型的一般策略。
