Details

Autor(en) / Beteiligte

• Viéitez, Cristina
• Busby, Bede P.
• Ochoa, David
• Mateus, André
• Memon, Danish
• Galardini, Marco
• Yildiz, Umut
• Trovato, Matteo
• Jawed, Areeb
• Geiger, Alexander G.
• Oborská-Oplová, 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

Titel

High-throughput functional characterization of protein phosphorylation sites in yeast

Ist Teil von

• Nature biotechnology, 2022-03, Vol.40 (3), p.382-390

Ort / Verlag

New York: Nature Publishing Group US

Erscheinungsjahr

2022

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.