德国欧洲分子生物学实验室Pedro Beltrao、Athanasios Typas和Mikhail M. Savitski研究组合作揭示酵母中蛋白质磷酸化位点的高通量功能特性。该项研究成果发表在2021年10月18日出版的《自然—生物技术》上。
在这项研究中，他们设计了一种化学遗传方法来研究酿酒酵母中磷酸化位点的功能相关性。他们构建了474 个在特定磷酸化位点发生突变的酵母菌株，这些菌株连同一个基因缺失文库在 102 种条件下进行了适应性筛选。在这些磷酸化位点中，42% 表现出生长表型，表明这些更有可能是功能性的。他们根据其生长曲线与基因缺失的相似性推断了它们的功能，并通过热蛋白质组分析和脂质组学验证了一个亚群。
高比例表现出在相应基因缺失中未见的表型，表明功能获得效应。对于人类中保守的磷酸化位点，酵母表型的严重程度表明它们与人类功能相关。这种高通量方法允许大规模表征单个磷酸化位点。
据介绍，磷酸化是一种关键的翻译后修饰，涉及几乎所有细胞过程的调节。然而，在最近发现的数千个磷酸化位点中，只有不到 5% 得到了功能注释。
附：英文原文
Title: High-throughput functional characterization of protein phosphorylation sites in yeast
Author: Viitez, Cristina, Busby, Bede P., Ochoa, David, Mateus, Andr, Memon, Danish, Galardini, Marco, Yildiz, Umut, Trovato, Matteo, Jawed, Areeb, Geiger, Alexander G., Oborsk-Oplov, Michaela, Potel, Clement M., Vonesch, Sibylle C., Szu Tu, Chelsea, Shahraz, Mohammed, Stein, Frank, Steinmetz, Lars M., Panse, Vikram G., Noh, Kyung-Min, Savitski, Mikhail M., Typas, Athanasios, Beltrao, Pedro
Issue&Volume: 2021-10-18
Abstract: Phosphorylation is a critical post-translational modification involved in the regulation of almost all cellular processes. However, fewer than 5% of thousands of recently discovered phosphosites have been functionally annotated. In this study, we devised a chemical genetic approach to study the functional relevance of phosphosites in Saccharomyces cerevisiae. We generated 474 yeast strains with mutations in specific phosphosites that were screened for fitness in 102 conditions, along with a gene deletion library. Of these phosphosites, 42% exhibited growth phenotypes, suggesting that these are more likely functional. We inferred their function based on the similarity of their growth profiles with that of gene deletions and validated a subset by thermal proteome profiling and lipidomics. A high fraction exhibited phenotypes not seen in the corresponding gene deletion, suggestive of a gain-of-function effect. For phosphosites conserved in humans, the severity of the yeast phenotypes is indicative of their human functional relevance. This high-throughput approach allows for functionally characterizing individual phosphosites at scale. Phenotypic profiling of a phospho-deficient mutant library informs functional annotations of phosphosites.
DOI: 10.1038/s41587-021-01051-x
Source: https://www.nature.com/articles/s41587-021-01051-x
Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex
本期文章：《自然—生物技术》：Online/在线发表
