Role of Host Type IA Phosphoinositide 3-Kinase Pathway Components in Invasin-Mediated Internalization of Yersinia enterocolitica

Infect Immun. 2016 May 24;84(6):1826-1841. doi: 10.1128/IAI.00142-16. Print 2016 Jun.

Abstract

Many bacterial pathogens subvert mammalian type IA phosphoinositide 3-kinase (PI3K) in order to induce their internalization into host cells. How PI3K promotes internalization is not well understood. Also unclear is whether type IA PI3K affects different pathogens through similar or distinct mechanisms. Here, we performed an RNA interference (RNAi)-based screen to identify components of the type IA PI3K pathway involved in invasin-mediated entry of Yersinia enterocolitica, an enteropathogen that causes enteritis and lymphadenitis. The 69 genes targeted encode known upstream regulators or downstream effectors of PI3K. A similar RNAi screen was previously performed with the food-borne bacterium Listeria monocytogenes The results of the screen with Y. enterocolitica indicate that at least nine members of the PI3K pathway are needed for invasin-mediated entry. Several of these proteins, including centaurin-α1, Dock180, focal adhesion kinase (FAK), Grp1, LL5α, LL5β, and PLD2 (phospholipase D2), were recruited to sites of entry. In addition, centaurin-α1, FAK, PLD2, and mTOR were required for remodeling of the actin cytoskeleton during entry. Six of the human proteins affecting invasin-dependent internalization also promote InlB-mediated entry of L. monocytogenes Our results identify several host proteins that mediate invasin-induced effects on the actin cytoskeleton and indicate that a subset of PI3K pathway components promote internalization of both Y. enterocolitica and L. monocytogenes.

MeSH terms

  • Actin Cytoskeleton / metabolism*
  • Actin Cytoskeleton / microbiology
  • Actin Cytoskeleton / ultrastructure
  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism
  • Adhesins, Bacterial / genetics*
  • Adhesins, Bacterial / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Class Ia Phosphatidylinositol 3-Kinase / genetics*
  • Class Ia Phosphatidylinositol 3-Kinase / metabolism
  • Focal Adhesion Kinase 1 / genetics
  • Focal Adhesion Kinase 1 / metabolism
  • Gene Expression Regulation
  • HeLa Cells
  • Host-Pathogen Interactions*
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Listeria monocytogenes / genetics*
  • Listeria monocytogenes / growth & development
  • Listeria monocytogenes / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Phosphoinositide-3 Kinase Inhibitors
  • Phospholipase D / genetics
  • Phospholipase D / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Signal Transduction
  • TOR Serine-Threonine Kinases / genetics
  • TOR Serine-Threonine Kinases / metabolism
  • Yersinia enterocolitica / genetics*
  • Yersinia enterocolitica / growth & development
  • Yersinia enterocolitica / metabolism
  • rac GTP-Binding Proteins / genetics
  • rac GTP-Binding Proteins / metabolism

Substances

  • ADAP1 protein, human
  • Adaptor Proteins, Signal Transducing
  • Adhesins, Bacterial
  • Carrier Proteins
  • DOCK1 protein, human
  • Intracellular Signaling Peptides and Proteins
  • Nerve Tissue Proteins
  • PHLDB1 protein, human
  • PHLDB2 protein, human
  • Phosphoinositide-3 Kinase Inhibitors
  • RNA, Small Interfering
  • Receptors, Cytoplasmic and Nuclear
  • phosphatidylinositol receptors
  • invasin, Yersinia
  • MTOR protein, human
  • TOR Serine-Threonine Kinases
  • Class Ia Phosphatidylinositol 3-Kinase
  • Focal Adhesion Kinase 1
  • PTK2 protein, human
  • phospholipase D2
  • Phospholipase D
  • rac GTP-Binding Proteins