Integrin-based diffusion barrier separates membrane domains enabling the formation of microbiostatic frustrated phagosomes

Elife. 2018 Mar 19:7:e34798. doi: 10.7554/eLife.34798.

Abstract

Candida albicans hyphae can reach enormous lengths, precluding their internalization by phagocytes. Nevertheless, macrophages engulf a portion of the hypha, generating incompletely sealed tubular phagosomes. These frustrated phagosomes are stabilized by a thick cuff of F-actin that polymerizes in response to non-canonical activation of integrins by fungal glycan. Despite their continuity, the surface and invaginating phagosomal membranes retain a strikingly distinct lipid composition. PtdIns(4,5)P2 is present at the plasmalemma but is not detectable in the phagosomal membrane, while PtdIns(3)P and PtdIns(3,4,5)P3 co-exist in the phagosomes yet are absent from the surface membrane. Moreover, endo-lysosomal proteins are present only in the phagosomal membrane. Fluorescence recovery after photobleaching revealed the presence of a diffusion barrier that maintains the identity of the open tubular phagosome separate from the plasmalemma. Formation of this barrier depends on Syk, Pyk2/Fak and formin-dependent actin assembly. Antimicrobial mechanisms can thereby be deployed, limiting the growth of the hyphae.

Keywords: Candida albicans; cell biology; integrins; macrophage; phagocytosis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antifungal Agents / pharmacology
  • Candida albicans / physiology
  • Cell Membrane / chemistry
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Female
  • Humans
  • Hyphae / physiology
  • Hyphae / ultrastructure
  • Integrins / metabolism*
  • Macrophages / drug effects
  • Macrophages / microbiology
  • Macrophages / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Phagocytosis / drug effects
  • Phagocytosis / physiology*
  • Phagosomes / drug effects
  • Phagosomes / metabolism*
  • Phagosomes / microbiology
  • Phosphatidylinositol 4,5-Diphosphate / metabolism
  • Phosphatidylinositol Phosphates / metabolism
  • RAW 264.7 Cells

Substances

  • Antifungal Agents
  • Integrins
  • Phosphatidylinositol 4,5-Diphosphate
  • Phosphatidylinositol Phosphates
  • phosphatidylinositol 3,4,5-triphosphate