title: The Dictyostelium prestalk inducer differentiation-inducing factor-1 (DIF-1) triggers unexpectedly complex global phosphorylation changes.
abstract: Differentiation-inducing factor-1 (DIF-1) is a polyketide that induces Dictyostelium amoebae to differentiate as prestalk cells. We performed a global quantitative screen for phosphorylation changes that occur within the first minutes after addition of DIF-1, using a triple-label SILAC approach. This revealed a new world of DIF-1-controlled signaling, with changes in components of the MAPK and protein kinase B signaling pathways, components of the actinomyosin cytoskeletal signaling networks, and a broad range of small GTPases and their regulators. The results also provide evidence that the Ca(2+)/calmodulin-dependent phosphatase calcineurin plays a role in DIF-1 signaling to the DimB prestalk transcription factor. At the global level, DIF-1 causes a major shift in the phosphorylation/dephosphorylation equilibrium toward net dephosphorylation. Of interest, many of the sites that are dephosphorylated in response to DIF-1 are phosphorylated in response to extracellular cAMP signaling. This accords with studies that suggest an antagonism between the two inducers and also with the rapid dephosphorylation of the cAMP receptor that we observe in response to DIF-1 and with the known inhibitory effect of DIF-1 on chemotaxis to cAMP. All MS data are available via ProteomeXchange with identifier PXD001555.
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Sugden
authors.affiliation: Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster LA1 4YG, United Kingdom.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Urbaniak
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Araki
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom J.G.Williams@dundee.ac.uk.
authors.forename: Jeffrey G
authors.initials: JG
authors.lastname: Williams
mesh_terms: D000595:Amino Acid Sequence; D019703:Calcineurin / Q000737:chemistry; D002633:Chemotaxis; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000378:metabolism*; D006588:Hexanones / Q000378:metabolism*; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D010766:Phosphorylation; D061065:Polyketides / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C490570:Differentiation-inducing factor-1, Dictyostelium; D006588:Hexanones; D006843:Hydrocarbons, Chlorinated; D061065:Polyketides; D015800:Protozoan Proteins; D000242:Cyclic AMP; D019703:Calcineurin