The yeast p24 complex regulates GPI-anchored protein transport and quality control by monitoring anchor remodeling

Castillon, Guillaume A.; Aguilera-Romero, Auxiliadora; Manzano-López, Javier; Epstein, Sharon; Kajiwara, Kentaro; Funato, Kouichi; Watanabe, Reika; Riezman, Howard; Muñiz, Manuel

doi:10.1091/mbc.e11-04-0294

Cited by 120 publications

(168 citation statements)

References 49 publications

(80 reference statements)

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“…The XbaI-XhoI fragments containing OSH4-36HA from FKP161 and HA-hCERT from FKP171 were also subcloned into XbaI-XhoI sites of pRS416GPD to obtain pRS416GPD-OSH4-36HA (FKP353) and pRS416GPD-HA-hCERT (FKP353), respectively. A LEU2 plasmid expressing Sec13p-mCherry (YCplac111-Sec13-mCh) was constructed as described previously (Castillon et al, 2011), by PCR using the primers 59-gggatatcaggaggcttccgagattttgg-39 and 59-gggatatcctcgcaggtctgcagcgaggcgcc-39 followed by enzymatic digestion with EcoRV and ligation. To generate LEU2 plasmids expressing wild-type Scs2 (pRS415-SCS2), mutant scs2 K[80/84/119/122]A (pRS415-scs2-K[80/84/119/122]A) and L84D/L86A (pRS415-scs2-L84D/L86A) under its own promoter, the fragments containing wild-type SCS2 from YCplac33-SCS2 (pKY151) (Kagiwada et al, 1998) and mutant scs2 from YCplac33-mutant scs2 vectors, respectively, which were constructed as described previously (Kagiwada and Hashimoto, 2007), were subcloned into HindIII-BamHI sites of pRS415.…”

Section: Strains and Plasmidsmentioning

confidence: 99%

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Osh proteins regulate COPII-mediated vesicular transport of ceramide from the endoplasmic reticulum in budding yeast

Kajiwara

Ikeda

Aguilera-Romero

et al. 2013

Journal of Cell Science

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Lipids synthesized at the endoplasmic reticulum (ER) are delivered to the Golgi by vesicular and non-vesicular pathways. ER-to-Golgi transport is crucial for maintaining the different membrane lipid composition and identities of organelles. Despite their importance, mechanisms regulating transport remain elusive. Here we report that in yeast coat protein complex II (COPII) vesicle-mediated transport of ceramide from the ER to the Golgi requires oxysterol-binding protein homologs, Osh proteins, which have been implicated in lipid homeostasis. Because Osh proteins are not required to transport proteins to the Golgi, these results indicate a specific requirement for the Osh proteins in the transport of ceramide. In addition, we provide evidence that Osh proteins play a negative role in COPII vesicle biogenesis. Together, our data suggest that ceramide transport and sphingolipid levels between the ER and Golgi are maintained by two distinct functions of Osh proteins, which negatively regulate COPII vesicle formation and positively control a later stage, presumably fusion of ceramide-enriched vesicles with Golgi compartments.

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Section: Strains and Plasmidsmentioning

confidence: 99%

“…proteins are packaged into budding COPII vesicles, cells were transformed with YCplac111-Sec13-mCh and were observed using a AXIOZ1 microscope as described previously (Castillon et al, 2011).…”

Section: Strains and Plasmidsmentioning

confidence: 99%

Osh proteins regulate COPII-mediated vesicular transport of ceramide from the endoplasmic reticulum in budding yeast

Kajiwara

Ikeda

Aguilera-Romero

et al. 2013

Journal of Cell Science

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“…VI , VII). In yeast, it has been shown that a defect in GPI anchor remodeling or traffi cking results in ER stress and induces the upregulation of the unfolded protein response (UPR) (40)(41)(42). We assessed ER stress by measuring transcriptional activation of the UPR.…”

Section: Very Long Chain Sphingomyelin Levels Are Partially Affected mentioning

confidence: 99%

Glycosylphosphatidylinositol anchors regulate glycosphingolipid levels

Loizides‐Mangold

David

Nesatyy

et al. 2012

Journal of Lipid Research

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This article is available online at http://www.jlr.org anchor has the core structure phosphatidylinositol (PI)-glucosamine (GlcN)-(Mannose) 3 -phosphoethanolamine (EtN-P), which is conserved among all species. After biosynthesis, the GPI anchor is attached posttranslationally to the newly generated C terminus of certain eukaryotic proteins destined for anchoring thereby tethering the protein to the membrane surface by the glycolipid moiety. GPI-anchored proteins can be released from the cell surface by phosphatidylinositol specifi c phospholipases and this cleavage event can induce major conformational changes on the GPIanchored protein itself ( 2 ).At least three organelles, the endoplasmic reticulum (ER), Golgi, and peroxisomes, are involved in the biosynthesis and remodeling of the GPI anchor. The biosynthesis is initiated on the outer side of the ER membrane. After the fi rst two reactions, the GPI anchor precursor is fl ipped and biosynthesis continues on the luminal side of the ER where the diacyl chains of phosphatidylinositol are then replaced by alkyl-acyl chains. This step is impaired in mutants of the peroxisomal alkyl phospholipid biosynthesis pathway ( 3 ).After protein attachment, the GPI anchor undergoes complex remodeling that begins in the ER with the removal of the inositol-linked acyl chain ( 4 ) and the remodeling of the GPI glycan part (5). Glycan remodeling is crucial for sorting GPI-anchored proteins into ER exit sites and their subsequent ER to Golgi transport ( 6 ). In mammalian cells, remodeling of the GPI anchor is then continued in the Golgi where the unsaturated fatty acid of the GPI anchor is replaced by a saturated fatty acid chain ( 7 ). Abstract Glycosylphosphatidylinositol (GPI) anchor biosynthesis takes place in the endoplasmic reticulum (ER).After protein attachment, the GPI anchor is transported to the Golgi where it undergoes fatty acid remodeling. The ER exit of GPI-anchored proteins is controlled by glycan remodeling and p24 complexes act as cargo receptors for GPI anchor sorting into COPII vesicles. In this study, we have characterized the lipid profi le of mammalian cell lines that have a defect in GPI anchor biosynthesis. Depending on which step of GPI anchor biosynthesis the cells were defective, we observed sphingolipid changes predominantly for very long chain monoglycosylated ceramides (HexCer). We found that the structure of the GPI anchor plays an important role in the control of HexCer levels. GPI anchordefi cient cells that generate short truncated GPI anchor intermediates showed a decrease in very long chain HexCer levels. Cells that synthesize GPI anchors but have a defect in GPI anchor remodeling in the ER have a general increase in HexCer levels. GPI-transamidase-defi cient cells that produce no GPI-anchored proteins but generate complete free GPI anchors had unchanged levels of HexCer. In contrast, sphingomyelin levels were mostly unaffected. We therefore propose a model in which the transport of very long chain ceramide from the ER to Golgi is regulated by...

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“…Results are presented as arbitrary units detected because absolute amounts could not be determined without an identical internal standard. The mass spectrometry yeast, emp24 and erv25 for instance, have been shown to induce the unfolded protein response (UPR) (23)(24)(25).…”

Section: Esi-ms Analysismentioning

confidence: 99%

Activation of the unfolded protein response pathway causes ceramide accumulation in yeast and INS-1E insulinoma cells

Epstein

Kirkpatrick

Castillon

et al. 2012

Journal of Lipid Research

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The yeast p24 complex regulates GPI-anchored protein transport and quality control by monitoring anchor remodeling

Cited by 120 publications

References 49 publications

Osh proteins regulate COPII-mediated vesicular transport of ceramide from the endoplasmic reticulum in budding yeast

Osh proteins regulate COPII-mediated vesicular transport of ceramide from the endoplasmic reticulum in budding yeast

Glycosylphosphatidylinositol anchors regulate glycosphingolipid levels

Activation of the unfolded protein response pathway causes ceramide accumulation in yeast and INS-1E insulinoma cells

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