Genetic evidence that an endosymbiont-derived endoplasmic reticulum-associated protein degradation (ERAD) system functions in import of apicoplast proteins

J Biol Chem. 2009 Nov 27;284(48):33683-91. doi: 10.1074/jbc.M109.044024. Epub 2009 Oct 6.

Abstract

Most apicomplexan parasites harbor a relict chloroplast, the apicoplast, that is critical for their survival. Whereas the apicoplast maintains a small genome, the bulk of its proteins are nuclear encoded and imported into the organelle. Several models have been proposed to explain how proteins might cross the four membranes that surround the apicoplast; however, experimental data discriminating these models are largely missing. Here we present genetic evidence that apicoplast protein import depends on elements derived from the ER-associated protein degradation (ERAD) system of the endosymbiont. We identified two sets of ERAD components in Toxoplasma gondii, one associated with the ER and cytoplasm and one localized to the membranes of the apicoplast. We engineered a conditional null mutant in apicoplast Der1, the putative pore of the apicoplast ERAD complex, and found that loss of Der1(Ap) results in loss of apicoplast protein import and subsequent death of the parasite.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / classification
  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism
  • Animals
  • Blotting, Western
  • Cell Cycle Proteins / classification
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cryoelectron Microscopy
  • Endoplasmic Reticulum / metabolism*
  • Eukaryota / physiology
  • Fibroblasts / parasitology
  • Fibroblasts / ultrastructure
  • Fluorescent Antibody Technique
  • Humans
  • Intracellular Membranes / metabolism
  • Molecular Sequence Data
  • Mutation
  • Phylogeny
  • Plastids / metabolism*
  • Protein Transport
  • Protozoan Proteins / classification
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Sequence Analysis, DNA
  • Symbiosis
  • Toxoplasma / genetics
  • Toxoplasma / metabolism*
  • Toxoplasma / microbiology
  • Valosin Containing Protein

Substances

  • Cell Cycle Proteins
  • Protozoan Proteins
  • Adenosine Triphosphatases
  • Valosin Containing Protein

Associated data

  • GENBANK/EJ976520
  • GENBANK/FJ976516
  • GENBANK/FJ976517
  • GENBANK/FJ976518
  • GENBANK/FJ976519
  • GENBANK/FJ976521
  • GENBANK/FJ976522