title: Three antagonistic cyclic di-GMP-catabolizing enzymes promote differential Dot/Icm effector delivery and intracellular survival at the early steps of Legionella pneumophila infection.
abstract: Legionella pneumophila is an intracellular pathogen which replicates within protozoan cells and can accidently infect alveolar macrophages, causing an acute pneumonia in humans. The second messenger cyclic di-GMP (c-di-GMP) has been shown to play key roles in the regulation of various bacterial processes, including virulence. While investigating the function of the 22 potential c-di-GMP-metabolizing enzymes of the L. pneumophila Lens strain, we found three that directly contribute to its ability to infect both protozoan and mammalian cells. These three enzymes display diguanylate cyclase (Lpl0780), phosphodiesterase (Lpl1118), and bifunctional diguanylate cyclase/phosphodiesterase (Lpl0922) activities, which are all required for the survival and intracellular replication of L. pneumophila. Mutants with deletions of the corresponding genes are efficiently taken up by phagocytic cells but are partially defective for the escape of the Legionella-containing vacuole (LCV) from the host degradative endocytic pathway and result in lower survival. In addition, Lpl1118 is required for efficient endoplasmic reticulum recruitment to the LCV. Trafficking and biogenesis of the LCV are dependent upon the orchestrated actions of several type 4 secretion system Dot/Icm effectors proteins, which exhibit differentially altered translocation in the three mutants. While translocation of some effectors remained unchanged, others appeared over- and undertranslocated. A general translocation offset of the large repertoire of Dot/Icm effectors may be responsible for the observed defects in the trafficking and biogenesis of the LCV. Our results suggest that L. pneumophila uses cyclic di-GMP signaling to fine-tune effector delivery and ensure effective evasion of the host degradative pathways and establishment of a replicative vacuole.
authors.affiliation: International Center for Infectiology Research (CIRI), Legionella Pathogenesis Group, Université de Lyon, Lyon, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Allombert
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Lazzaroni
authors.forename: Nathalie
authors.initials: N
authors.lastname: Baïlo
authors.forename: Christophe
authors.initials: C
authors.lastname: Gilbert
authors.forename: Xavier
authors.initials: X
authors.lastname: Charpentier
authors.forename: Patricia
authors.initials: P
authors.lastname: Doublet
authors.forename: Anne
authors.initials: A
authors.lastname: Vianney
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D006801:Humans; D016952:Legionella pneumophila / Q000378:metabolism*; D007877:Legionnaires' Disease / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D010586:Phagocytes / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D019761:Phosphorus-Oxygen Lyases / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology; D020298:U937 Cells; D014774:Virulence / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D010727:Phosphoric Diester Hydrolases; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase; D006152:Cyclic GMP