title: PrkC-mediated phosphorylation of overexpressed YvcK protein regulates PBP1 protein localization in Bacillus subtilis mreB mutant cells.
abstract: The YvcK protein has been shown to be necessary for growth under gluconeogenic conditions in Bacillus subtilis. Amazingly, its overproduction rescues growth and morphology defects of the actin-like protein MreB deletion mutant by restoration of PBP1 localization. In this work, we observed that YvcK was phosphorylated at Thr-304 by the protein kinase PrkC and that phosphorylated YvcK was dephosphorylated by the cognate phosphatase PrpC. We show that neither substitution of this threonine with a constitutively phosphorylated mimicking glutamic acid residue or a phosphorylation-dead mimicking alanine residue nor deletion of prkC or prpC altered the ability of B. subtilis to grow under gluconeogenic conditions. However, we observed that a prpC mutant and a yvcK mutant were more sensitive to bacitracin compared with the WT strain. In addition, the bacitracin sensitivity of strains in which YvcK Thr-304 was replaced with either an alanine or a glutamic acid residue was also affected. We also analyzed rescue of the mreB mutant strain by overproduction of YvcK in which the phosphorylation site was substituted. We show that YvcK T304A overproduction did not rescue the mreB mutant aberrant morphology due to PBP1 mislocalization. The same observation was made in an mreB prkC double mutant overproducing YvcK. Altogether, these data show that YvcK may have two distinct functions: 1) in carbon source utilization independent of its phosphorylation level and 2) in cell wall biosynthesis and morphogenesis through its phosphorylation state.
authors.affiliation: From the Laboratoire de Chimie Bactérienne, UMR 7283, Institut de Microbiologie de la Méditerranée, CNRS, Aix-Marseille Université, 13009 Marseille and.
authors.forename: Elodie
authors.initials: E
authors.lastname: Foulquier
authors.affiliation: From the Laboratoire de Chimie Bactérienne, UMR 7283, Institut de Microbiologie de la Méditerranée, CNRS, Aix-Marseille Université, 13009 Marseille and.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Pompeo
authors.affiliation: From the Laboratoire de Chimie Bactérienne, UMR 7283, Institut de Microbiologie de la Méditerranée, CNRS, Aix-Marseille Université, 13009 Marseille and the Bases Moléculaires et Structurales des Systèmes Infectieux, UMR 5086, CNRS, Université de Lyon, 69367 Lyon Cedex 07, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Freton
authors.affiliation: From the Laboratoire de Chimie Bactérienne, UMR 7283, Institut de Microbiologie de la Méditerranée, CNRS, Aix-Marseille Université, 13009 Marseille and.
authors.forename: Baptiste
authors.initials: B
authors.lastname: Cordier
authors.affiliation: the Bases Moléculaires et Structurales des Systèmes Infectieux, UMR 5086, CNRS, Université de Lyon, 69367 Lyon Cedex 07, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Grangeasse
authors.affiliation: From the Laboratoire de Chimie Bactérienne, UMR 7283, Institut de Microbiologie de la Méditerranée, CNRS, Aix-Marseille Université, 13009 Marseille and galinier@imm.cnrs.fr.
authors.forename: Anne
authors.initials: A
authors.lastname: Galinier
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001414:Bacitracin / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D015153:Blotting, Western; D024881:Drug Resistance, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D053719:Tandem Mass Spectrometry; D013912:Threonine / Q000378:metabolism; D020798:Two-Hybrid System Techniques
chemical_list: D001426:Bacterial Proteins; D001414:Bacitracin; D013912:Threonine
keywords: Antibiotic Resistance; Bacillus; Cell Wall; Gluconeogenesis; Morphogenesis; Protein Phosphorylation