title: Functional characterization of the Mycobacterium tuberculosis serine/threonine kinase PknJ.
abstract: Eukaryotic-like Ser/Thr protein kinases (STPKs) are present in many bacterial species, where they control various physiological and virulence processes by enabling microbial adaptation to specific environmental signals. PknJ is the only member of the 11 STPKs identified in Mycobacterium tuberculosis that still awaits characterization. Here we report that PknJ is a functional kinase that forms dimers in vitro, and contains a single transmembrane domain. Using a high-density peptide-chip-based technology, multiple potential mycobacterial targets were identified for PknJ. We confirmed PknJ-dependent phosphorylation of four of these targets: PknJ itself, which autophosphorylates at Thr(168), Thr(171) and Thr(173) residues; the transcriptional regulator EmbR; the methyltransferase MmaA4/Hma involved in mycolic acid biosynthesis; and the dipeptidase PepE, whose encoding gene is located next to pknJ in the mycobacterial genome. Our results provide a number of candidate phospho-targets for PknJ and possibly other mycobacterial STPKs that could be studied to investigate the role of STPKs in M. tuberculosis physiology and virulence.
authors.affiliation: Inserm Equipe Avenir Biology of Intracellular Pathogens, Institut Pasteur Korea, Seoul, Republic of Korea.|Unit of Mycobacterial Genetics, Institut Pasteur, Paris, France.
authors.forename: Jichan
authors.initials: J
authors.lastname: Jang
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Stella
authors.affiliation: Unit of Mycobacterial Genetics, Institut Pasteur, Paris, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Boudou
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Levillain
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Eliette
authors.initials: E
authors.lastname: Darthuy
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Vaubourgeix
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Chongzhen
authors.initials: C
authors.lastname: Wang
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Bardou
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Germain
authors.initials: G
authors.lastname: Puzo
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Gilleron
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Burlet-Schiltz
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Monsarrat
authors.affiliation: Inserm Equipe Avenir Biology of Intracellular Pathogens, Institut Pasteur Korea, Seoul, Republic of Korea.
authors.forename: Priscille
authors.initials: P
authors.lastname: Brodin
authors.affiliation: Unit of Mycobacterial Genetics, Institut Pasteur, Paris, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gicquel
authors.affiliation: Université de Toulouse, Université Paul Sabatier, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France.|Unit of Mycobacterial Genetics, Institut Pasteur, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Neyrolles
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019281:Dimerization; D051379:Mice; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D013912:Threonine / Q000378:metabolism; D014376:Tuberculosis / Q000382:microbiology; D014774:Virulence
chemical_list: D013912:Threonine; D012694:Serine; D017346:Protein Serine-Threonine Kinases