A genomewide overexpression screen identifies genes involved in the phosphatidylinositol 3-kinase pathway in the human protozoan parasite Entamoeba histolytica.

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  • Author(s): Koushik AB;Koushik AB; Welter BH; Rock ML; Temesvari LA
  • Source:
    Eukaryotic cell [Eukaryot Cell] 2014 Mar; Vol. 13 (3), pp. 401-11. Date of Electronic Publication: 2014 Jan 17.
  • Publication Type:
    Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101130731 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1535-9786 (Electronic) Linking ISSN: 15359786 NLM ISO Abbreviation: Eukaryot Cell Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : American Society for Microbiology, c2002-
    • Subject Terms:
      Genome, Protozoan*; Entamoeba histolytica/*genetics ; Phosphatidylinositol 3-Kinase/*metabolism ; Signal Transduction/*genetics; Amino Acid Sequence ; Chromones/pharmacology ; Entamoeba histolytica/metabolism ; Microfilament Proteins/genetics ; Microfilament Proteins/metabolism ; Molecular Sequence Data ; Morpholines/pharmacology ; Phosphatidylinositol 3-Kinase/genetics ; Phosphoinositide-3 Kinase Inhibitors ; Protein Kinases/genetics ; Protein Kinases/metabolism ; Protozoan Proteins/genetics ; Protozoan Proteins/metabolism
    • Abstract:
      Entamoeba histolytica is a protozoan parasite that causes amoebic dysentery and liver abscess. E. histolytica relies on motility, phagocytosis, host cell adhesion, and proteolysis of extracellular matrix for virulence. In eukaryotic cells, these processes are mediated in part by phosphatidylinositol 3-kinase (PI3K) signaling. Thus, PI3K may be critical for virulence. We utilized a functional genomics approach to identify genes whose products may operate in the PI3K pathway in E. histolytica. We treated a population of trophozoites that were overexpressing genes from a cDNA library with a near-lethal dose of the PI3K inhibitor wortmannin. This screen was based on the rationale that survivors would be overexpressing gene products that directly or indirectly function in the PI3K pathway. We sequenced the overexpressed genes in survivors and identified a cDNA encoding a Rap GTPase, a protein previously shown to participate in the PI3K pathway. This supports the validity of our approach. Genes encoding a coactosin-like protein, EhCoactosin, and a serine-rich E. histolytica protein (SREHP) were also identified. Cells overexpressing EhCoactosin or SREHP were also less sensitive to a second PI3K inhibitor, LY294002. This corroborates the link between these proteins and PI3K. Finally, a mutant cell line with an increased level of phosphatidylinositol (3,4,5)-triphosphate, the product of PI3K activity, exhibited increased expression of SREHP and EhCoactosin. This further supports the functional connection between these proteins and PI3K in E. histolytica. To our knowledge, this is the first forward-genetics screen adapted to reveal genes participating in a signal transduction pathway in this pathogen.
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    • Grant Information:
      P20 GM103444 United States GM NIGMS NIH HHS; R21 AI081100 United States AI NIAID NIH HHS; R21 AI081100-01 United States AI NIAID NIH HHS
    • Accession Number:
      0 (Chromones)
      0 (Microfilament Proteins)
      0 (Morpholines)
      0 (Phosphoinositide-3 Kinase Inhibitors)
      0 (Protozoan Proteins)
      31M2U1DVID (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one)
      EC 2.7.- (Protein Kinases)
      EC 2.7.1.- (RAP kinase)
      EC 2.7.1.137 (Phosphatidylinositol 3-Kinase)
    • Publication Date:
      Date Created: 20140121 Date Completed: 20141125 Latest Revision: 20240520
    • Publication Date:
      20240520
    • Accession Number:
      PMC3957588
    • Accession Number:
      10.1128/EC.00329-13
    • Accession Number:
      24442890