Protein phosphorylation evolution across 18 fungal species.
For each species whole cell extracts were digested separately with trypsin and LysC. Each digest was IMAC enriched and analyzed by LC-MS/MS in technical triplicate on an LTQ Orbitrap Velos using a 120min method. Data was searched against the corresponding species using Sequest/Comet, and filtered to 1% FDR at the PSM level.
For S.Cerevisiae, S.Pombe and S.Mikatae, cells were grown in triplicate under 3 different conditions: YPD, YPD + mild temperature stress and optimized "best media". For these cells whole cell extracts were produced, digested with trypsin and IMAC enriched and analyzed by LC-MS/MS on an LTQ Orbitrap Velos using a 120min method. Data was searched against the corresponding species using Sequest/Comet, and filtered to 1% FDR at the PSM level.
S. cerevisiae cells transformed with tagged versions of CDC33 WT, alanine (S28A) and glutamic acid (S28E) mutants were generated. Mutant strain were grown three distinct (heavy, medium and light) SILAC media under overexpression conditions. Cells for each construct were harvested and pooled together and used for the pull down experiments, done in triplicate. Pulldown eluate was digested with LysC and analyzed on a QExactive instrument with 90min method. Data was searched against S.Cerevisiae database including the tagged CDC33 proteins using Sequest/Comet, and filtered to 1% FDR at the PSM level.
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: evolution ; phosphorylation ; yeast ; phosphoproteomics
Principal Investigators: (in alphabetical order) |
Judit Vill�n |
Submitting User: | jvillen |
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Romain A. Studer, Ricard A. Rodriguez-Mias, Kelsey M. Haas, Joanne I. Hsu, Cristina Vieitez, Carme Sole, Danielle L. Swaney, Lindsay B. Stanford, Ivan Liachko, Rene Bottcher, Maitreya J. Dunham, Eulalia de Nadal, Francesc Posas, Pedro Beltrao, Judit Villen.
Evolution of protein phosphorylation across 18 fungal species.
Science. 2016 Oct 14;354(6309):229-232. doi: 10.1126/science.aaf2144.
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