title: Temporal dynamics of the Saccharopolyspora erythraea phosphoproteome.
abstract: Actinomycetes undergo a dramatic reorganization of metabolic and cellular machinery during a brief period of growth arrest ("metabolic switch") preceding mycelia differentiation and the onset of secondary metabolite biosynthesis. This study explores the role of phosphorylation in coordinating the metabolic switch in the industrial actinomycete Saccharopolyspora erythraea. A total of 109 phosphopeptides from 88 proteins were detected across a 150-h fermentation using open-profile two-dimensional LC-MS proteomics and TiO(2) enrichment. Quantitative analysis of the phosphopeptides and their unphosphorylated cognates was possible for 20 pairs that also displayed constant total protein expression. Enzymes from central carbon metabolism such as putative acetyl-coenzyme A carboxylase, isocitrate lyase, and 2-oxoglutarate dehydrogenase changed dramatically in the degree of phosphorylation during the stationary phase, suggesting metabolic rearrangement for the reutilization of substrates and the production of polyketide precursors. In addition, an enzyme involved in cellular response to environmental stress, trypsin-like serine protease (SACE_6340/NC_009142_6216), decreased in phosphorylation during the growth arrest stage. More important, enzymes related to the regulation of protein synthesis underwent rapid phosphorylation changes during this stage. Whereas the degree of phosphorylation of ribonuclease Rne/Rng (SACE_1406/NC_009142_1388) increased during the metabolic switch, that of two ribosomal proteins, S6 (SACE_7351/NC_009142_7233) and S32 (SACE_6101/NC_009142_5981), dramatically decreased during this stage of the fermentation, supporting the hypothesis that ribosome subpopulations differentially regulate translation before and after the metabolic switch. Overall, we show the great potential of phosphoproteomic studies to explain microbial physiology and specifically provide evidence of dynamic protein phosphorylation events across the developmental cycle of actinomycetes.
authors.affiliation: §Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, St Lucia, QLD 4072, Australia.
authors.forename: Cuauhtemoc
authors.initials: C
authors.lastname: Licona-Cassani
authors.forename: Sooa
authors.initials: S
authors.lastname: Lim
authors.forename: Esteban
authors.initials: E
authors.lastname: Marcellin
authors.forename: Lars K
authors.initials: LK
authors.lastname: Nielsen
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D013058:Mass Spectrometry; D010748:Phosphopeptides / Q000378:metabolism; D010766:Phosphorylation; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D016930:Saccharopolyspora / Q000254:growth & development* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010748:Phosphopeptides; D020543:Proteome