title: Nucleolin, a shuttle protein promoting infection of human monocytes by Francisella tularensis.
abstract: BACKGROUND
  Francisella tularensis is a highly virulent facultative intracellular bacterium, disseminating in vivo mainly within host mononuclear phagocytes. After entry into macrophages, F. tularensis initially resides in a phagosomal compartment, whose maturation is then arrested. Bacteria escape rapidly into the cytoplasm, where they replicate freely. We recently demonstrated that nucleolin, an eukaryotic protein able to traffic from the nucleus to the cell surface, acted as a surface receptor for F. tularensis LVS on human monocyte-like THP-1 cells.
  
  
  RESULTS
  Here, we followed the fate of nucleolin once F. tularensis has been endocytosed. We first confirmed by siRNA silencing experiments that expression of nucleolin protein was essential for binding of LVS on human macrophage-type THP-1 cells. We then showed that nucleolin co-localized with intracellular bacteria in the phagosomal compartment. Strikingly, in that compartment, nucleolin also co-localized with LAMP-1, a late endosomal marker. Co-immunoprecipation assays further demonstrated an interaction of nucleolin with LAMP-1. Co-localization of nucleolin with LVS was no longer detectable at 24 h when bacteria were multiplying in the cytoplasm. In contrast, with an iglC mutant of LVS, which remains trapped into the phagosomal compartment, or with inert particles, nucleolin/bacteria co-localization remained almost constant.
  
  
  CONCLUSIONS
  We herein confirm the importance of nucleolin expression for LVS binding and its specificity as nucleolin is not involved in binding of another intracellular pathogen as L. monocytogenes or an inert particle. Association of nucleolin with F. tularensis during infection continues intracellularly after endocytosis of the bacteria. The present work therefore unravels for the first time the presence of nucleolin in the phagosomal compartment of macrophages.
authors.affiliation: Université Paris Descartes, Faculté de Médecine Necker Enfants-Malades, Paris, France. monique.barel@inserm.fr
authors.forename: Monique
authors.initials: M
authors.lastname: Barel
authors.forename: Karin
authors.initials: K
authors.lastname: Meibom
authors.forename: Alain
authors.initials: A
authors.lastname: Charbit
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004072:Digitonin / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D005604:Francisella tularensis / Q000378:metabolism*; D006801:Humans; D052118:Lysosomal-Associated Membrane Protein 1 / Q000378:metabolism; D009000:Monocytes / Q000166:cytology / Q000382:microbiology*; D009154:Mutation; D010587:Phagocytosis; D010750:Phosphoproteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012503:Saponins / Q000378:metabolism; D014406:Tularemia / Q000378:metabolism*; D000097796:Nucleolin
chemical_list: D004072:Digitonin; D052118:Lysosomal-Associated Membrane Protein 1; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D012503:Saponins; D000097796:Nucleolin