Phosphoinositides differentially regulate bacterial uptake and Nramp1-induced resistance to Legionella infection in Dictyostelium

J Cell Sci. 2010 Dec 1;123(Pt 23):4039-51. doi: 10.1242/jcs.072124. Epub 2010 Nov 2.

Abstract

Membrane phosphatidylinositides recruit cytosolic proteins to regulate phagocytosis, macropinocytosis and endolysosomal vesicle maturation. Here, we describe effects of inactivation of PI3K, PTEN or PLC on Escherichia coli and Legionella pneumophila uptake by the professional phagocyte Dictyostelium discoideum. We show that L. pneumophila is engulfed by macropinocytosis, a process that is partially sensitive to PI3K inactivation, unlike phagocytosis of E. coli. Both processes are blocked by PLC inhibition. Whereas E. coli is rapidly digested, Legionella proliferates intracellularly. Proliferation is blocked by constitutively expressing Nramp1, an endolysosomal iron transporter that confers resistance against invasive bacteria. Inactivation of PI3K, but not PTEN or PLC, enhances Legionella infection and suppresses the protective effect of Nramp1 overexpression. PI3K activity is restricted to early infection and is not mediated by effects on the actin cytoskeleton; rather L. pneumophila, in contrast to E. coli, subverts phosphoinositide-sensitive fusion of Legionella-containing macropinosomes with acidic vesicles, without affecting Nramp1 recruitment. A model is presented to explain how Legionella escapes fusion with acidic vesicles and Nramp1-induced resistance to pathogens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cation Transport Proteins / genetics
  • Cation Transport Proteins / metabolism*
  • Dictyostelium / enzymology
  • Dictyostelium / genetics
  • Dictyostelium / microbiology*
  • Dictyostelium / physiology*
  • Escherichia coli / physiology
  • Legionella pneumophila / physiology*
  • Phagocytosis*
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphatidylinositols / metabolism*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Type C Phospholipases / genetics
  • Type C Phospholipases / metabolism

Substances

  • Cation Transport Proteins
  • Phosphatidylinositols
  • Protozoan Proteins
  • natural resistance-associated macrophage protein 1
  • Type C Phospholipases