Glycosylphosphatidylinositol-specific phospholipase C regulates transferrin endocytosis in the African trypanosome

Sandesh Subramanya, C Frank Hardin, Dietmar Steverding, Kojo Mensa-Wilmot

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

GPI-PLC (glycosylphosphatidylinositol-specific phospholipase C) is expressed in bloodstream-form Trypanosoma brucei, a protozoan that causes human African trypanosomiasis. Loss of genes encoding GPI-PLC reduces the virulence of a pleomorphic strain of the parasite, for reasons that are not clear. In the present paper, we report that GPI-PLC stimulates endocytosis of transferrin by 300-500%. Surprisingly, GPI-PLC is not detected at endosomes, suggesting that the enzyme does not interact directly with the endosomal machinery. We therefore hypothesized that a diffusible product of the GPI-PLC enzyme reaction [possibly DAG (diacylglycerol)] mediated the biological effects of the protein. Two sets of data support this assertion. First, a catalytically inactive Q81L mutant of GPI-PLC, expressed in a GPI-PLC-null background, had no effect on endocytosis, indicating that enzyme activity is essential for the protein to stimulate endocytosis. Secondly, the exogenous DAGs OAG (1-oleyl-2-acetyl-sn-glycerol) and DMG (dimyristoylglycerol) independently stimulated endocytosis of transferrin. Furthermore, the DAG mimic PMA, a phorbol ester, also activated endocytosis in T. brucei. DAG-stimulated endocytosis is a novel pathway in the trypanosome. We surmise that (i) GPI-PLC regulates transferrin endocytosis in T. brucei, (ii) GPI-PLC is a signalling enzyme, and (iii) DAG is a second messenger for GPI-PLC. We propose that regulation of endocytosis is a physiological function of GPI-PLC in bloodstream T. brucei.
Original languageEnglish
Pages (from-to)685-694
Number of pages10
JournalBiochemical Journal
Volume417
Issue number3
DOIs
Publication statusPublished - 2009

Keywords

  • Animals
  • Diglycerides
  • Endocytosis
  • Glycosylphosphatidylinositol Diacylglycerol-Lyase
  • Microscopy, Fluorescence
  • Phorbol Esters
  • Protozoan Proteins
  • Transferrin
  • Trypanosoma brucei brucei

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