Phospholipase D activity couples plasma membrane endocytosis with retromer dependent recycling

Elife. 2016 Nov 16;5:e18515. doi: 10.7554/eLife.18515.

Abstract

During illumination, the light-sensitive plasma membrane (rhabdomere) of Drosophila photoreceptors undergoes turnover with consequent changes in size and composition. However, the mechanism by which illumination is coupled to rhabdomere turnover remains unclear. We find that photoreceptors contain a light-dependent phospholipase D (PLD) activity. During illumination, loss of PLD resulted in an enhanced reduction in rhabdomere size, accumulation of Rab7 positive, rhodopsin1-containing vesicles (RLVs) in the cell body and reduced rhodopsin protein. These phenotypes were associated with reduced levels of phosphatidic acid, the product of PLD activity and were rescued by reconstitution with catalytically active PLD. In wild-type photoreceptors, during illumination, enhanced PLD activity was sufficient to clear RLVs from the cell body by a process dependent on Arf1-GTP levels and retromer complex function. Thus, during illumination, PLD activity couples endocytosis of RLVs with their recycling to the plasma membrane thus maintaining plasma membrane size and composition.

Keywords: Arf1; D. melanogaster; biochemistry; cell biology; lipid signalling; phosphatidic acid; phospholipase D; retromer; rhodopsin.

MeSH terms

  • ADP-Ribosylation Factor 1 / genetics
  • ADP-Ribosylation Factor 1 / metabolism
  • Animals
  • Cell Membrane / metabolism*
  • Cell Membrane / radiation effects
  • Cell Membrane / ultrastructure
  • Cytoplasmic Vesicles / metabolism
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / enzymology*
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / radiation effects
  • Endocytosis / radiation effects*
  • Gene Expression
  • Genetic Complementation Test
  • Guanosine Triphosphate / metabolism
  • Light
  • Phosphatidic Acids / metabolism
  • Phospholipase D / genetics
  • Phospholipase D / metabolism*
  • Photic Stimulation
  • Photoreceptor Cells, Invertebrate / metabolism*
  • Photoreceptor Cells, Invertebrate / radiation effects
  • Photoreceptor Cells, Invertebrate / ultrastructure
  • Rhodopsin / genetics
  • Rhodopsin / metabolism
  • Vision, Ocular / physiology
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / metabolism
  • rab7 GTP-Binding Proteins

Substances

  • Drosophila Proteins
  • Phosphatidic Acids
  • rab7 GTP-Binding Proteins
  • Guanosine Triphosphate
  • Rhodopsin
  • Phospholipase D
  • ADP-Ribosylation Factor 1
  • rab GTP-Binding Proteins