Mammalian Erv46 localizes to the endoplasmic reticulum–Golgi intermediate compartment and to cis-Golgi cisternae

Orci, Lelio; Ravazzola, Mariella; Mack, G; Barlowe, Charles; Otte, Stefan

doi:10.1073/pnas.0730885100

Cited by 21 publications

(17 citation statements)

References 33 publications

(44 reference statements)

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“…We first compared the localization of ERGIC-32 and hErv46 by confocal immunofluorescence microscopy. Mouse Erv46 localizes to the cis-Golgi and the ERGIC (41). Using the rabbit anti-mouse Erv46 antibody, which cross-reacts with the human antigen, we found prominent colocalization of C-terminally HA-tagged ERGIC-32 with hErv46 in the Golgi area.…”

Section: Purification Of Ergic Membranes From Bfa-treated Cells-mentioning

confidence: 92%

Proteomics of Endoplasmic Reticulum-Golgi Intermediate Compartment (ERGIC) Membranes from Brefeldin A-treated HepG2 Cells Identifies ERGIC-32, a New Cycling Protein That Interacts with Human Erv46

Breuza

Halbeisen

Jenö

et al. 2004

Journal of Biological Chemistry

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113

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Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. Membranes enriched 110-fold over the homogenate for ERGIC-53 were obtained and analyzed by mass spectrometry. Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. ERGIC-32 localizes to the ERGIC and partially colocalizes with the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi. ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. We propose that ERGIC-32 functions as a modulator of the hErv41-hErv46 complex by stabilizing hErv46. Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway.

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Section: Purification Of Ergic Membranes From Bfa-treated Cells-mentioning

confidence: 92%

Proteomics of Endoplasmic Reticulum-Golgi Intermediate Compartment (ERGIC) Membranes from Brefeldin A-treated HepG2 Cells Identifies ERGIC-32, a New Cycling Protein That Interacts with Human Erv46

Breuza

Halbeisen

Jenö

et al. 2004

Journal of Biological Chemistry

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113

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“…According to real-time RT-PCR analysis, DDR1 , HSP90B1 , SDC1 , RPSA , ERGIC3 , and LPCAT1 were up-regulated significantly in NSCLCs, while GPX3 , TIMP3 were down-regulated significantly. ERGIC3 is located in endoplasmic reticulum and Golgi apparatus of NRK cells [11], however, the function of ERGIC3 is unclear in lung cancer. Therefore, expression of ERGIC3 in NSCLCs was further confirmed at the protein level by western blot and immunohistochemistry analysis and we studied the pathophysiological functions of ERGIC 3.…”

Section: Introductionmentioning

confidence: 99%

Suppression subtractive hybridization identified differentially expressed genes in lung adenocarcinoma: ERGIC3as a novel lung cancer-related gene

Huang

et al. 2013

BMC Cancer

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Background: To understand the carcinogenesis caused by accumulated genetic and epigenetic alterations and seek novel biomarkers for various cancers, studying differentially expressed genes between cancerous and normal tissues is crucial. In the study, two cDNA libraries of lung cancer were constructed and screened for identification of differentially expressed genes. Methods: Two cDNA libraries of differentially expressed genes were constructed using lung adenocarcinoma tissue and adjacent nonmalignant lung tissue by suppression subtractive hybridization. The data of the cDNA libraries were then analyzed and compared using bioinformatics analysis. Levels of mRNA and protein were measured by quantitative real-time polymerase chain reaction (q-RT-PCR) and western blot respectively, as well as expression and localization of proteins were determined by immunostaining. Gene functions were investigated using proliferation and migration assays after gene silencing and gene over-expression. Results: Two libraries of differentially expressed genes were obtained. The forward-subtracted library (FSL) and the reverse-subtracted library (RSL) contained 177 and 59 genes, respectively. Bioinformatic analysis demonstrated that these genes were involved in a wide range of cellular functions. The vast majority of these genes were newly identified to be abnormally expressed in lung cancer. In the first stage of the screening for 16 genes, we compared lung cancer tissues with their adjacent non-malignant tissues at the mRNA level, and found six genes (ERGIC3, DDR1, HSP90B1, SDC1, RPSA, and LPCAT1) from the FSL were significantly up-regulated while two genes (GPX3 and TIMP3) from the RSL were significantly down-regulated (P < 0.05). The ERGIC3 protein was also over-expressed in lung cancer tissues and cultured cells, and expression of ERGIC3 was correlated with the differentiated degree and histological type of lung cancer. The up-regulation of ERGIC3 could promote cellular migration and proliferation in vitro.

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“…These include the localization of the Arf-GEF GBF1 (Zhao et al 2002;Garcia-Mata et al 2003) and ER vesicle protein 46 (Orci et al 2003) to cis-Golgi compartments and Golgin GCC88 to trans-Golgi compartments (Luke et al 2003). In one recent study, a wide range of constructs derived from Arf1 was scored for localization to cis or trans compartments within the Golgi apparatus using light microscopy .…”

mentioning

confidence: 99%

Confocal Microscopy-based Linescan Methodologies for Intra-Golgi Localization of Proteins

Dejgaard

Murshid

Dee

et al. 2007

J Histochem Cytochem.

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Localization of resident Golgi proteins to earlier ( cis) or later ( trans) Golgi compartments has traditionally required quantitative immunocytochemistry and electron microscopy, which are inaccessible to many researchers. For this reason, light microscopy has often been used, initially for localization of Golgi glycotransferases and, more recently, for other Golgi proteins (e.g., Arf1, GBF1, Rab6). Quantitation of light microscopic intra-Golgi localization can be problematic. We describe here a novel quantitative light microscopic methodology using linescans crossing the Golgi ribbon. Our method determines a localization for the unknown protein in a one-dimensional coordinate system in which 0.0 corresponds to localization of a cis marker and 1.0 to localization of a trans marker. We also describe a variant of this methodology in which Golgi morphology is simplified by nocodazole-induced dispersal into ministacks, allowing a fully automated analysis. In our assay, β1,4-galactosyltransferase-YFP and Golgin97 localize similarly to trans markers, whereas p115, GBF1, and p58-YFP are similarly near other cis markers. The medial Golgi protein α1,3–1,6-mannosidase II gives an intermediate localization in this assay. These methodologies may prove useful in instances where electron microscopy is technically difficult as well as when rapid analysis of large numbers of samples is required.

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Mammalian Erv46 localizes to the endoplasmic reticulum–Golgi intermediate compartment and to cis-Golgi cisternae

Cited by 21 publications

References 33 publications

Proteomics of Endoplasmic Reticulum-Golgi Intermediate Compartment (ERGIC) Membranes from Brefeldin A-treated HepG2 Cells Identifies ERGIC-32, a New Cycling Protein That Interacts with Human Erv46

Proteomics of Endoplasmic Reticulum-Golgi Intermediate Compartment (ERGIC) Membranes from Brefeldin A-treated HepG2 Cells Identifies ERGIC-32, a New Cycling Protein That Interacts with Human Erv46

Suppression subtractive hybridization identified differentially expressed genes in lung adenocarcinoma: ERGIC3as a novel lung cancer-related gene

Confocal Microscopy-based Linescan Methodologies for Intra-Golgi Localization of Proteins

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