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

Monnat, J; Neuhaus, E M; Pop, M S; Ferrari, D M; Kramer, B; Soldati, Thierry

doi:10.1091/mbc.11.10.3469

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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 Soldati, Thierry
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 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
Identifiers		DOI: 10.1091/mbc.11.10.3469 PMID: 11029049
Full text		Article (Published version) (1 MB) - 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. doi: 10.1091/mbc.11.10.3469 https://archive-ouverte.unige.ch/unige:18920