Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin a suggests an ER recycling pathway

Lippincott‐Schwartz, Jennifer; Donaldson, Julie G.; Schweizer, Anja; Berger, Eric G.; Hauri, Hans‐Peter; Yuan, Lydia C.; Klausner, Richard D.

doi:10.1016/0092-8674(90)90096-w

Cited by 949 publications

(678 citation statements)

References 49 publications

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“…might exist also within the ER, possibly as a consequence of the differential diffusion of the molecules recyded the level of the cis-Golgi (Lippincott-Schwartz et al, 1990;Haugejorden et al, 1991;Lewis and Pelham, 1992). Data on both CSQ and calreticulin (Thomas et al, 1989;Peter et al, 1992) document the existence of additional pathways of membrane recyding, originated at various levels of the GC.…”

Section: Specializations Of the Er Luminal Contentmentioning

confidence: 99%

Endoplasmic reticulum: a dynamic patchwork of specialized subregions.

Sitia

Meldolesi

1992

MBoC

150

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Section: Specializations Of the Er Luminal Contentmentioning

confidence: 99%

Endoplasmic reticulum: a dynamic patchwork of specialized subregions.

Sitia

Meldolesi

1992

MBoC

150

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“…In particular, ceramide coupled to the fluorophore boron dipyrromethene difluoride (BODIPY, or C5-DMB-Cer) was shown to be bright, resistant to photobleaching, and to stain the Golgi complex more efficiently than other cellular membranes. In this work we show that the transport of C5-DMB-Cer to the cell surface and its subsequent loss from the cells is a time-dependent process that is blocked by Brefeldin A (BFA), a drug that also blocks secretion (Misumi et al, 1986;Lippincott-Schwartz et al, 1990;Hendricks et al, 1992). In addition, cells with known thermosensitive defects in secretion retain the fluorescent label longer than wild-type cells at the restrictive temperature.…”

Section: Introductionmentioning

confidence: 99%

A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.

Ktistakis

Kao

Wang

et al. 1995

MBoC

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The use of reporter proteins to study the regulation of secretion has often been complicated by posttranslational processing events that influence the secretion of certain proteins, but are not part of the cellular mechanisms that specifically regulate secretion. This has been a particular limitation for the isolation of mammalian secretion mutants, which has typically been a slow process. To provide a reporter of secretory activity independent of protein processing events, cells were labeled with the fluorescent lipid analogue C5-DMB-ceramide (ceramide coupled to the fluorophore boron dipyrromethene difluoride) and its secretion was followed by fluorescence microscopy and fluorescenceactivated cell sorting. Brefeldin A, which severely inhibits secretion in Chinese hamster ovary cells, blocked secretion of C5-DMB-ceramide. At high temperature, export of C5-DMB-ceramide was inhibited in HRP-1 cells, which have a conditional defect in secretion. Using C5-DMB-ceramide as a reporter of secretory activity, several different pulse-chase protocols were designed that selected mutant Chinese hamster ovary cells that were resistant to the drug brefeldin A and others that were defective in the transport of glycoproteins to the cell surface. Mutant cells of either type were identified in a mutagenized population at a frequency of 10-6. Thus, the fluorescent lipid C5-DMBceramide can be used as a specific marker of secretory activity, providing an efficient, general approach for isolating mammalian cells with defects in the secretory pathway.

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“…The two proteins are differently distributed in the endomembrane system. ERGIC-53 is associated with the ERGolgi intermediate compartment (ERGIC; Hauri and Schweizer, 1992) and continuously recycles between the endoplasmic reticulum (ER) and the ERGIC (Lippincott-Schwartz et al, 1990;Foguet, Clausen, Tang, Hong, and Hauri, unpublished data), while VIP36 is presumed to recycle between the Golgi apparatus and the plasma membrane . Together these features of ERGIC-53 and VIP36 would fit the predictions for intracellular sorting receptors.…”

Section: Introductionmentioning

confidence: 99%

ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins.

Itin

Roche

Monsigny

et al. 1996

MBoC

167

148

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Based on sequence homologies with leguminous lectins, the intermediate compartment marker proposed to be a member of a putative new class of animal lectins associated with the secretory pathway. Independently, a promyelocytic protein, MR60, was purified by mannose-column chromatography, and a cDNA was isolated that matched MR60 peptide sequences. This cDNA was identical to that of and homologies with the animal lectin family of the galectins were noticed. Not all peptide sequences of MR60, however, were found in ERGIC-53, raising the possibility that another protein associated with ERGIC-53 may possess the lectin activity. Here, we provide the first direct evidence for a lectin function of ERGIC-53. Overexpressed ERGIC-53 binds to a mannose column in a calcium-dependent manner and also co-stains with mannosylated neoglycoprotein in a morphological binding assay. By using a sequential elution protocol we show that ERGIC-53 has selectivity for mannose and low affinity for glucose and GlcNAc, but no affinity for galactose. To experimentally address the putative homology of ERGIC-53 to leguminous lectins, a highly conserved protein family with an invariant asparagine essential for carbohydrate binding, we substituted the corresponding asparagine in ERGIC-53. This mutation, as well as a mutation affecting a second site in the putative carbohydrate recognition domain, abolished mannosecolumn binding and co-staining with mannosylated neoglycoprotein. These findings establish ERGIC-53 as a lectin and provide functional evidence for its relationship to leguminous lectins. Based on its monosaccharide specificity, domain organization, and recycling properties, we propose ERGIC-53 to function as a sorting receptor for glycoproteins in the early secretory pathway.

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Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin a suggests an ER recycling pathway

Cited by 949 publications

References 49 publications

Endoplasmic reticulum: a dynamic patchwork of specialized subregions.

Endoplasmic reticulum: a dynamic patchwork of specialized subregions.

A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.

ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins.

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