Scientific article
Open access

Identification of a novel saturable endoplasmic reticulum localization mechanism mediated by the C-terminus of a Dictyostelium protein disulfide isomerase

Published inMolecular biology of the cell, vol. 11, no. 10, p. 3469-3484
Publication date2000

Localization of soluble endoplasmic reticulum (ER) resident proteins is likely achieved by the complementary action of retrieval and retention mechanisms. Whereas the machinery involving the H/KDEL and related retrieval signals in targeting escapees back to the ER is well characterized, other mechanisms including retention are still poorly understood. We have identified a protein disulfide isomerase (Dd-PDI) lacking the HDEL retrieval signal normally found at the C terminus of ER residents in Dictyostelium discoideum. Here we demonstrate that its 57 residue C-terminal domain is necessary for intracellular retention of Dd-PDI and sufficient to localize a green fluorescent protein (GFP) chimera to the ER, especially to the nuclear envelope. Dd-PDI and GFP-PDI57 are recovered in similar cation-dependent complexes. The overexpression of GFP-PDI57 leads to disruption of endogenous PDI complexes and induces the secretion of PDI, whereas overexpression of a GFP-HDEL chimera induces the secretion of endogenous calreticulin, revealing the presence of two independent and saturable mechanisms. Finally, low-level expression of Dd-PDI but not of PDI truncated of its 57 C-terminal residues complements the otherwise lethal yeast TRG1/PDI1 null mutation, demonstrating functional disulfide isomerase activity and ER localization. Altogether, these results indicate that the PDI57 peptide contains ER localization determinants recognized by a conserved machinery present in D. discoideum and Saccharomyces cerevisiae.

  • Amino Acid Sequence
  • Animals
  • Cycloheximide/pharmacology
  • Dictyostelium/enzymology/physiology/ultrastructure
  • Endoplasmic Reticulum/physiology/ultrastructure
  • Genes, Reporter
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins/genetics
  • Molecular Sequence Data
  • Oligopeptides
  • Protein Disulfide-Isomerases/chemistry/genetics/metabolism
  • Protein Sorting Signals
  • Protein Transport
  • Rats
  • Recombinant Fusion Proteins/chemistry/metabolism
  • Sequence Alignment
  • Sequence Deletion
  • Sequence Homology, Amino Acid
  • Signal Transduction
Affiliation Not a UNIGE publication
Citation (ISO format)
MONNAT, J et al. Identification of a novel saturable endoplasmic reticulum localization mechanism mediated by the C-terminus of a Dictyostelium protein disulfide isomerase. In: Molecular biology of the cell, 2000, vol. 11, n° 10, p. 3469–3484. doi: 10.1091/mbc.11.10.3469
Main files (1)
Article (Published version)
ISSN of the journal1059-1524

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