Adaptor protein complex 1 mediates the transport of lysosomal proteins from a Golgi-like organelle to peripheral vacuoles in the primitive eukaryote Giardia lamblia

Mol Biol Cell. 2004 Jul;15(7):3053-60. doi: 10.1091/mbc.e03-10-0744. Epub 2004 Apr 23.

Abstract

Giardia lamblia is an early branching protist that possesses peripheral vacuoles (PVs) with characteristics of lysosome-like organelles, located underneath the plasma membrane. In more evolved cells, lysosomal protein trafficking is achieved by cargo recognition involving adaptor protein (AP) complexes that recognize specific amino acid sequences (tyrosine and/or dileucine motifs) within the cytoplasmic tail of membrane proteins. Previously, we reported that Giardia has a tyrosine-based sorting system, which mediates the targeting of a membrane-associated cysteine protease (encystation-specific cysteine protease, ESCP) to the PVs. Here, we show that Giardia AP1 mediates the transport of ESCP and the soluble acid phosphatase (AcPh) to the PVs. By using the yeast two-hybrid assay we found that the ESCP tyrosine-based motif interacts specifically with the medium subunit of AP1 (Gimicroa). Hemagglutinin-tagged Gimicroa colocalizes with ESCP and AcPh and coimmunoprecipitates with clathrin, suggesting that protein trafficking toward the PVs is clathrin-adaptin dependent. Targeted disruption of Gimicroa results in mislocalization of ESCP and AcPh but not of variant-specific surface proteins. Our results suggest that, unlike mammalian cells, only AP1 is involved in anterograde protein trafficking to the PVs in Giardia. Moreover, even though Giardia trophozoites lack a morphologically discernible Golgi apparatus, the presence of a clathrin-adaptor system suggests that this parasite possess a primitive secretory organelle capable of sorting proteins similar to that of more evolved cells.

MeSH terms

  • Acid Phosphatase / analysis
  • Acid Phosphatase / metabolism
  • Adaptor Protein Complex 1 / analysis
  • Adaptor Protein Complex 1 / physiology*
  • Amino Acid Motifs
  • Animals
  • Brefeldin A / pharmacology
  • Clathrin / metabolism
  • Cysteine Endopeptidases / analysis
  • Cysteine Endopeptidases / metabolism
  • Giardia lamblia / immunology
  • Giardia lamblia / physiology*
  • Giardia lamblia / ultrastructure
  • Golgi Apparatus / physiology*
  • Protein Transport / drug effects
  • Proteins / analysis
  • Proteins / metabolism*
  • Vacuoles / immunology
  • Vacuoles / physiology*

Substances

  • Adaptor Protein Complex 1
  • Clathrin
  • Proteins
  • lysosomal proteins
  • Brefeldin A
  • Acid Phosphatase
  • Cysteine Endopeptidases