当前位置: X-MOL 学术Nat. Biotechnol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
High-throughput functional characterization of protein phosphorylation sites in yeast
Nature Biotechnology ( IF 46.9 ) Pub Date : 2021-10-18 , DOI: 10.1038/s41587-021-01051-x
Cristina Viéitez 1, 2 , Bede P Busby 1, 2 , David Ochoa 1 , André Mateus 2 , Danish Memon 1 , Marco Galardini 1 , Umut Yildiz 2, 3 , Matteo Trovato 2, 3 , Areeb Jawed 2 , Alexander G Geiger 4 , Michaela Oborská-Oplová 4, 5 , Clement M Potel 2 , Sibylle C Vonesch 2 , Chelsea Szu Tu 2 , Mohammed Shahraz 1 , Frank Stein 2 , Lars M Steinmetz 2, 6, 7 , Vikram G Panse 4, 8 , Kyung-Min Noh 2 , Mikhail M Savitski 2 , Athanasios Typas 2 , Pedro Beltrao 1, 2
Affiliation  

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.



中文翻译:

酵母中蛋白质磷酸化位点的高通量功能表征

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位点中,只有不到 5% 的功能得到了注释。在这项研究中,我们设计了一种化学遗传学方法来研究酿酒酵母中磷酸盐的功能相关性. 我们生成了 474 种在特定磷酸位点发生突变的酵母菌株,这些菌株在 102 种条件下进行了适应性筛选,以及一个基因缺失文库。在这些磷酸盐位点中,42% 表现出生长表型,表明它们更有可能发挥功能。我们根据它们的生长曲线与基因缺失的相似性推断它们的功能,并通过热蛋白质组分析和脂质组学验证了一个子集。很大一部分表现出在相应基因缺失中未见的表型,这表明存在功能获得效应。对于在人类中保守的磷酸盐位点,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地对单个磷酸盐进行功能表征。

更新日期:2021-10-18
down
wechat
bug