Valenti Rosario, David Yotam, Edilbi Dunya, Dubreuil Benjamin, Boshnakovska Angela, Asraf Yeynit, Salame Tomer-Meir, Sass Ehud, Rehling Peter, Schuldiner Maya
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Department of Cellular Biochemistry, University Medical Center Göttingen, Göttingen, Germany.
J Cell Biol. 2025 Feb 3;224(2). doi: 10.1083/jcb.202409050. Epub 2024 Dec 18.
Genome-wide collections of yeast strains, known as libraries, revolutionized the way systematic studies are carried out. Specifically, libraries that involve a cellular perturbation, such as the deletion collection, have facilitated key biological discoveries. However, short-term rewiring and long-term accumulation of suppressor mutations often obscure the functional consequences of such perturbations. We present the AID library which supplies "on demand" protein depletion to overcome these limitations. Here, each protein is tagged with a green fluorescent protein (GFP) and an auxin-inducible degron (AID), enabling rapid protein depletion that can be quantified systematically using the GFP element. We characterized the degradation response of all strains and demonstrated its utility by revisiting seminal yeast screens for genes involved in cell cycle progression as well as mitochondrial distribution and morphology. In addition to recapitulating known phenotypes, we also uncovered proteins with previously unrecognized roles in these central processes. Hence, our tool expands our knowledge of cellular biology and physiology by enabling access to phenotypes that are central to cellular physiology and therefore rapidly equilibrated.
全基因组酵母菌株集合，即所谓的文库，彻底改变了系统研究的开展方式。具体而言，涉及细胞扰动的文库，如缺失文库，推动了关键生物学发现。然而，短期的重新布线和抑制突变的长期积累常常掩盖了此类扰动的功能后果。我们展示了AID文库，它提供“按需”蛋白质消耗以克服这些限制。在此，每种蛋白质都标记有绿色荧光蛋白（GFP）和生长素诱导降解结构域（AID），实现了快速的蛋白质消耗，可使用GFP元件进行系统定量。我们对所有菌株的降解反应进行了表征，并通过重新审视关于参与细胞周期进程以及线粒体分布和形态的基因的重要酵母筛选，证明了其效用。除了重现已知表型外，我们还发现了在这些核心过程中具有先前未被认识作用的蛋白质。因此，我们的工具通过能够获取对细胞生理学至关重要且因此快速平衡的表型，扩展了我们对细胞生物学和生理学的认识。
