Compartmental organization of Golgi-specific protein modification and vacuolar protein sorting events defined in a yeast sec18 (NSF) mutant.

Graham, Todd R.; Emr, Scott D.

doi:10.1083/jcb.114.2.207

Cited by 393 publications

(312 citation statements)

References 56 publications

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“…4A). A partially glycosylated precursor of the secreted protein accumulated in temperature sensitive sec18 cells (23,24), when ER exit is blocked at restrictive temperature (Fig. 4B).…”

Section: Resultsmentioning

confidence: 98%

Membrane association is a determinant for substrate recognition by PMT4 protein O -mannosyltransferases

Hutzler

Schmid

Bernard

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…4A). A partially glycosylated precursor of the secreted protein accumulated in temperature sensitive sec18 cells (23,24), when ER exit is blocked at restrictive temperature (Fig. 4B).…”

Section: Resultsmentioning

confidence: 98%

Membrane association is a determinant for substrate recognition by PMT4 protein O -mannosyltransferases

Hutzler

Schmid

Bernard

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…In the ER, it undergoes signal peptide cleavage and N-linked core-glycosylation (12 potential sites), generating an 80 kDa protein ( Figure 1A) that accumulates in the sec18, impaired primarily in ER-to-Golgi trafficking (Graham and Emr, 1991) (Figures 1B, 2). This protein undergoes outer chain glycosylation in the Golgi apparatus by the addition of long mannose chains of variable length, generating a mature protein (100-150 kDa) that is targeted via secretory vesicles to the periplasm.…”

Section: Resultsmentioning

confidence: 99%

Mutants defective in secretory/vacuolar pathways in the EUROFAN collection of yeast disruptants

Avaro

Belgareh‐Touzé

Sibella-Argüelles

et al. 2002

Yeast

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We have screened the EUROFAN (European Functional Analysis Network) deletion strain collection for yeast mutants defective in secretory/vacuolar pathways and/or associated biochemical modifications. We used systematic Western immunoblotting to analyse the electrophoretic pattern of several markers of the secretory/vacuolar pathways, the soluble a-factor, the periplasmic glycoprotein invertase, the plasma membrane GPIanchored protein Gas1p, and two vacuolar proteins, the soluble carboxypeptidase Y and the membrane-bound alkaline phosphatase, which are targeted to the vacuole by different pathways. We also used colony immunoblotting to monitor the secretion of carboxypeptidase Y into the medium, to identify disruptants impaired in vacuolar targeting. We identified 25 mutants among the 631 deletion strains. Nine of these mutants were disrupted in genes identified in recent years on the basis of their involvement in trafficking (VPS53, VAC7, VAM6, APM3, SYS1), or glycosylation (ALG12, ALG9, OST4, ROT2). Three of these genes were identified on the basis of trafficking defects by ourselves and others within the EUROFAN project (TLG2, RCY1, MON2). The deletion of ERV29, which encodes a COPII vesicle protein, impaired carboxypeptidase Y trafficking from the endoplasmic reticulum to the Golgi apparatus. We also identified eight unknown ORFs, the deletion of which reduced Golgi glycosylation or impaired the Golgi to vacuole trafficking of carboxypeptidase Y. YJR044c, which we identified as a new VPS gene, encodes a protein with numerous homologues of unknown function in sequence databases.

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“…The latter mechanism would lead to Golgi-specific limited outer-chain glycosyl modification of the core glycosylated mutant proteins. These outer-chain glycosyl modifications occur in at least two distinct Golgi compartments (Franzusoff and Schekman, 1989 ;Graham and Emr, 1991). In the first Golgi compartment, a-l-6-mannosyl residues are added to the core glycosyls.…”

Section: Pra* and Cpy* Are Not A-1+6 Or A-1-+3 Mannosylatedmentioning

confidence: 99%

Analysis of two mutated vacuolar proteins reveals a degradation pathway in the endoplasmic reticulum or a related compartment of yeast

Finger

Knop

Wolf

1993

European Journal of Biochemistry

194

174

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The fate of a mutant form of each of the two yeast vacuolar enzymes proteinase yscA (PrA) and carboxypeptidase yscY (CPY) has been investigated. Both mutant proteins are rapidly degraded after entering the secretory pathway. Mutant PrA is deleted in 37 amino acids spanning the processing site region of the PrA pro‐peptide. The mutant enzyme shows no activity towards maturation of itself or other vacuolar hydrolases, a function of wild‐type PrA. Mutant CPY carries an Arg instead of a Gly residue in a highly conserved region, two positions distant from the active‐site Ser. In contrast to wild‐type CPY, the mutant form was quickly degraded by trypsin in vitro, indicating an altered structure. Using antisera specific for α‐1→6 and α‐1→3 outer‐chain mannose linkages, no Golgi‐specific carbohydrate modification could be detected on either mutant protein. Subcellular fractionation studies located both mutant enzymes in the endoplasmic reticulum. Degradation kinetics of both proteins show the same characteristics, indicating similar degradation pathways. The degradation process was shown to be independent of a functional sec18 gene product and takes place before Golgi‐specific carbohydrate modifications occur. The proteasome, the major proteolytic activity of the cytoplasm, is not involved in this degradation event. All degradation characteristics of the two mutant proteins are consistent with a degradation process within the endoplasmic reticulum (‘ER degradation’).

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Compartmental organization of Golgi-specific protein modification and vacuolar protein sorting events defined in a yeast sec18 (NSF) mutant.

Abstract: Abstract. The sec18 and sec23 secretory

Cited by 393 publications

References 56 publications

Membrane association is a determinant for substrate recognition by PMT4 protein O -mannosyltransferases

Membrane association is a determinant for substrate recognition by PMT4 protein O -mannosyltransferases

Mutants defective in secretory/vacuolar pathways in the EUROFAN collection of yeast disruptants

Analysis of two mutated vacuolar proteins reveals a degradation pathway in the endoplasmic reticulum or a related compartment of yeast

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