Rab14 is critical for maintenance of Mycobacterium tuberculosis phagosome maturation arrest

EMBO J. 2006 Nov 15;25(22):5250-9. doi: 10.1038/sj.emboj.7601407. Epub 2006 Nov 2.

Abstract

Mycobacterium tuberculosis arrests phagosomal maturation in infected macrophage, and, apart from health significance, provides a superb model system to dissect the phagolysosomal biogenesis pathway. Here, we demonstrate a critical role for the small GTPase Rab14 in maintaining mycobacterial phagosome maturation block. Four-dimensional microscopy showed that phagosomes containing live mycobacteria accumulated Rab14 following phagocytosis. The recruitment of Rab14 had strong functional consequence, as a knockdown of endogenous Rab14 by siRNA or overexpression of Rab14 dominant-negative mutants (Rab14S25N and Rab14N125I) released the maturation block and allowed phagosomes harboring live mycobacteria to progress into phagolysosomes. Conversely, overexpression of the wild-type Rab14 and the constitutively active mutant Rab14Q70L prevented phagosomes with dead mycobacteria from undergoing default maturation into phagolysosomal organelles. Mechanistic studies demonstrated a role for Rab14 in stimulating organellar fusion between phagosomes and early endosomes but not with late endosomes. Rab14 enables mycobacterial phagosomes to maintain early endosomal characteristics and avoid late endosomal/lysosomal degradative components.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Line
  • Endosomes / physiology
  • Humans
  • Intracellular Membranes / physiology
  • Macrophages / microbiology*
  • Macrophages / physiology
  • Membrane Fusion
  • Mice
  • Mutation
  • Mycobacterium tuberculosis / metabolism
  • Mycobacterium tuberculosis / physiology*
  • Phagosomes / microbiology
  • Phagosomes / physiology*
  • RNA, Small Interfering / genetics
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / physiology*

Substances

  • RNA, Small Interfering
  • rab GTP-Binding Proteins