title: Comparative proteomic analysis of Streptomyces lividans Wild-Type and ppk mutant strains reveals the importance of storage lipids for antibiotic biosynthesis.
abstract: Streptomyces lividans TK24 is a strain that naturally produces antibiotics at low levels, but dramatic overproduction of antibiotics occurs upon interruption of the ppk gene. However, the role of the Ppk enzyme in relation to the regulation of antibiotic biosynthesis remains poorly understood. In order to gain a better understanding of the phenotype of the ppk mutant, the proteomes of the wild-type (wt) and ppk mutant strains, grown for 96 h on R2YE medium limited in phosphate, were analyzed. Intracellular proteins were separated on two-dimensional (2D) gels, spots were quantified, and those showing a 3-fold variation or more were identified by mass spectrometry. The expression of 12 proteins increased and that of 29 decreased in the ppk mutant strain. Our results suggested that storage lipid degradation rather than hexose catabolism was taking place in the mutant. In order to validate this hypothesis, the triacylglycerol contents of the wt and ppk mutant strains of S. lividans as well as that of Streptomyces coelicolor M145, a strain that produces antibiotics at high levels and is closely related to S. lividans, were assessed using electron microscopy and thin-layer chromatography. These studies highlighted the large difference in triacylglycerol contents of the three strains and confirmed the hypothetical link between storage lipid metabolism and antibiotic biosynthesis in Streptomyces.
authors.affiliation: Institut de Biochimie et Biophysique Moléculaire et Cellulaire, CNRS UMR 8619, Université Paris-Sud, Orsay, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Le Maréchal
authors.forename: Paulette
authors.initials: P
authors.lastname: Decottignies
authors.forename: Christophe H
authors.initials: CH
authors.lastname: Marchand
authors.forename: Jeril
authors.initials: J
authors.lastname: Degrouard
authors.forename: Danièle
authors.initials: D
authors.lastname: Jaillard
authors.forename: Thierry
authors.initials: T
authors.lastname: Dulermo
authors.forename: Marine
authors.initials: M
authors.lastname: Froissard
authors.forename: Aleksey
authors.initials: A
authors.lastname: Smirnov
authors.forename: Violaine
authors.initials: V
authors.lastname: Chapuis
authors.forename: Marie-Joelle
authors.initials: MJ
authors.lastname: Virolle
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional; D017353:Gene Deletion; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D048170:Streptomyces lividans / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D020543:Proteome; D017853:Phosphotransferases (Alcohol Group Acceptor)