title: Phosphoproteomic analysis of protein phosphorylation networks in Tetrahymena thermophila, a model single-celled organism.
abstract: Tetrahymena thermophila is a widely used unicellular eukaryotic model organism in biological research and contains more than 1000 protein kinases and phosphatases with specificity for Ser/Thr/Tyr residues. However, only a few dozen phosphorylation sites in T. thermophila are known, presenting a major obstacle to further understanding of the regulatory roles of reversible phosphorylation in this organism. In this study, we used high-accuracy mass-spectrometry-based proteomics to conduct global and site-specific phosphoproteome profiling of T. thermophila. In total, 1384 phosphopeptides and 2238 phosphorylation sites from 1008 T. thermophila proteins were identified through the combined use of peptide prefractionation, TiO2 enrichment, and two-dimensional LC-MS/MS analysis. The identified phosphoproteins are implicated in the regulation of various biological processes such as transport, gene expression, and mRNA metabolic process. Moreover, integrated analysis of the T. thermophila phosphoproteome and gene network revealed the potential biological functions of many previously unannotated proteins and predicted some putative kinase-substrate pairs. Our data provide the first global survey of phosphorylation in T. thermophila using a phosphoproteomic approach and suggest a wide-ranging regulatory scope of this modification. The provided dataset is a valuable resource for the future understanding of signaling pathways in this important model organism.
authors.affiliation: Key Laboratory of Aquatic Biodiversity and Conservation, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, Hubei 430072, China;
authors.forename: Miao
authors.initials: M
authors.lastname: Tian
authors.forename: Xiulan
authors.initials: X
authors.lastname: Chen
authors.forename: Qian
authors.initials: Q
authors.lastname: Xiong
authors.forename: Jie
authors.initials: J
authors.lastname: Xiong
authors.forename: Chuanle
authors.initials: C
authors.lastname: Xiao
authors.forename: Feng
authors.initials: F
authors.lastname: Ge
authors.forename: Fuquan
authors.initials: F
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Miao
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D008962:Models, Theoretical; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000378:metabolism; D060066:Protein Interaction Maps; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D016808:Tetrahymena thermophila / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D020543:Proteome; D010770:Phosphotransferases