Scientific Article
previous document  unige:18920  next document
add to browser collection
Title

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

Authors
Monnat, J
Neuhaus, E M
Pop, M S
Ferrari, D M
Kramer, B
Published in Molecular Biology of the Cell. 2000, vol. 11, no. 10, p. 3469-84
Abstract 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.
Keywords Amino Acid SequenceAnimalsCycloheximide/pharmacologyDictyostelium/enzymology/physiology/ultrastructureEndoplasmic Reticulum/physiology/ultrastructureGenes, ReporterGreen Fluorescent ProteinsHumansLuminescent Proteins/geneticsMolecular Sequence DataOligopeptidesProtein Disulfide-Isomerases/chemistry/genetics/metabolismProtein Sorting SignalsProtein TransportRatsRecombinant Fusion Proteins/chemistry/metabolismSequence AlignmentSequence DeletionSequence Homology, Amino AcidSignal Transduction
Identifiers
PMID: 11029049
Full text
Article (Published version) (1 MB) - public document Free access
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. 3469-84. https://archive-ouverte.unige.ch/unige:18920

208 hits

324 downloads

Update

Deposited on : 2012-03-20

Export document
Format :
Citation style :