Molecular cloning of a novel 97‐kd Golgi complex autoantigen associated with Sjögren's syndrome

Griffith, Kevin J.; Chan, Edward K. L.; Lung, Chien‐Cheng; Hamel, John C.; Guo, Xiaoying; Miyachi, Kiyomitsu; Fritzler, Marvin J.

doi:10.1002/art.1780400920

Cited by 157 publications

(95 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We sought the identity of these structures by using appropriate markers and reagents for cytoplasmic organelles and vesicles in colocalization reactions with the index human serum ( Figure 1). The staining obtained with the index human serum did not colocalize with the Golgi compartments, as determined by studies that used antibodies to golgin-97 ( Figure 1A; Griffith et al, 1997) and the marker of the ER/Golgi intermediate compartment p58 (Lahtinen et al, 1996;Figure 1B) or TGN38, a marker for the trans-Golgi network (TGN38; our unpublished results; Luzio et al, 1990). Brefeldin, a fungal toxin known to rapidly dissociate the Golgi complex (Orci et al, 1991) did not affect the morphology, size, or number of these structures (our unpublished results).…”

Section: Gw182 Is Localized To Cytoplasmic Specklesmentioning

confidence: 86%

“…Clone 5.1 and the full-length GW182 construct were used as templates for in vitro transcription and translation (TnT; Promega, Madison, WI) in the presence of [ 35 S]methionine (Fritzler et al, 1995;Griffith et al, 1997). TnT reactions were conducted at 30°C for 1.5-2 h. To confirm the presence of translation products, 2-to 5-l samples were fractionated by SDS-PAGE and analyzed by autoradiography.…”

Section: In Vitro Transcription/translation and Immunoprecipitationmentioning

confidence: 99%

“…For colocalization studies primary antibodies to the following proteins were used: golgin-97 (murine monoclonal cdf4; Griffith et al, 1997), p58 (murine monoclonal, a gift from Dr. Tom Hobman, University of Alberta), TGN38 (murine monoclonal; Affinity Bioreagents, Golden, CO), clathrin heavy chain (murine monoclonal; Transduction Laboratories, Lexington, KY), rab9 (murine monoclonal, Affinity Bioreagents, Golden, CO), PMP70 (murine monoclonal; Zymed, San Francisco, CA), LAMP1 (murine monoclonal; Drs. August and Hildreth, Pharmacology and Molecular Sciences, John Hopkins University School of Medicine, Baltimore, MD), EEA1 (rabbit polyclonal antibody; Selak et al, 1999), cullin-1 (rabbit polyclonal antibody; NeoMarkers, Fremont, CA), caveolin (rabbit polyclonal antibody, Transduction Laboratories), and p19 skp1 (rabbit polyclonal antibody; NeoMarkers).…”

Section: Indirect Immunofluorescencementioning

confidence: 99%

See 2 more Smart Citations

A Phosphorylated Cytoplasmic Autoantigen, GW182, Associates with a Unique Population of Human mRNAs within Novel Cytoplasmic Speckles

Eystathioy

Chan

Tenenbaum

et al. 2002

MBoC

Self Cite

330

313

View full text Add to dashboard Cite

A novel human cellular structure has been identified that contains a unique autoimmune antigen and multiple messenger RNAs. This complex was discovered using an autoimmune serum from a patient with motor and sensory neuropathy and contains a protein of 182 kDa. The gene and cDNA encoding the protein indicated an open reading frame with glycine-tryptophan (GW) repeats and a single RNA recognition motif. Both the patient's serum and a rabbit serum raised against the recombinant GW protein costained discrete cytoplasmic speckles designated as GW bodies (GWBs) that do not overlap with the Golgi complex, endosomes, lysosomes, or peroxisomes. The mRNAs associated with GW182 represent a clustered set of transcripts that are presumed to reside within the GW complexes. We propose that the GW ribonucleoprotein complex is involved in the posttranscriptional regulation of gene expression by sequestering a specific subset of gene transcripts involved in cell growth and homeostasis.

show abstract

Section: Gw182 Is Localized To Cytoplasmic Specklesmentioning

confidence: 86%

Section: In Vitro Transcription/translation and Immunoprecipitationmentioning

confidence: 99%

Section: Indirect Immunofluorescencementioning

confidence: 99%

See 1 more Smart Citation

A Phosphorylated Cytoplasmic Autoantigen, GW182, Associates with a Unique Population of Human mRNAs within Novel Cytoplasmic Speckles

Eystathioy

Chan

Tenenbaum

et al. 2002

MBoC

Self Cite

330

313

View full text Add to dashboard Cite

show abstract

“…There are four human TGN golgins, namely p230/golgin-245, golgin-97, GCC185 and GCC88 (8)(9)(10)(11)(12)(13), which all belong to a subfamily of golgins characterized by a C-terminal GRIP domain (14)(15)(16). Recruitment of both p230/ golgin-245 (p230) and golgin-97 to the TGN is mediated through an interaction with the small G protein Arl1 (17)(18)(19)(20); however, these two golgins are localized to distinct membrane domains of the TGN (21).…”

mentioning

confidence: 99%

A trans -Golgi network golgin is required for the regulated secretion of TNF in activated macrophages in vivo

Lieu

Lock

Hammond

et al. 2008

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

View full text Add to dashboard Cite

The transmembrane precursor of tumor necrosis factor-␣ (TNF) exits the trans-Golgi network (TGN) in tubular carriers for subsequent trafficking and delivery to the cell surface; however, the molecular machinery responsible for Golgi export is unknown. We previously reported that members of the TGN golgin family are associated with subdomains and tubules of the TGN. Here, we show that the TGN golgin, p230/golgin-245 (p230), is essential for intracellular trafficking and cell surface delivery of TNF in transfected HeLa cells and activated macrophages. Live-cell imaging revealed that TNF transport from the TGN is mediated selectively by tubules and carriers marked by p230. Significantly, LPS activation of macrophages resulted in a dramatic increase of p230-labeled tubules and carriers emerging from the TGN, indicating that macrophages up-regulate the transport pathway for TNF export. Depletion of p230 in LPS-stimulated macrophages reduced cell surface delivery of TNF by >10-fold compared with control cells. To determine whether p230 depletion blocked TNF secretion in vivo, we generated retrogenic mice expressing a microRNAvector to silence p230. Bone-marrow stem cells were transduced with recombinant retrovirus containing microRNA constructs and transplanted into irradiated recipients. LPS-activated peritoneal macrophages from p230 miRNA retrogenic mice were depleted of p230 and had dramatically reduced levels of cell surface TNF. Overall, these studies have identified p230 as a key regulator of TNF secretion and have shown that LPS activation of macrophages results in increased Golgi carriers for export. Also, we have demonstrated a previously undescribed approach to control cytokine secretion by the specific silencing of trafficking machinery. intracellular trafficking ͉ RNAi transgenic mice ͉ TNF secretion ͉ inflammation ͉ membrane transport T umor necrosis factor-␣ (TNF) is the main proinflammatory cytokine made and secreted by inflammatory macrophages. Early release of TNF in response to LPS or other inflammatory signals enhances the activation and recruitment of T cells and ensures robust innate and acquired immune responses (1). The excessive secretion of TNF is also a prevalent and clinically significant problem in acute inflammation and in chronic inflammatory disease (2). Anti-TNF treatments have shown success in the treatment of rheumatoid arthritis, inflammatory bowel disease, and other conditions (3). Now improved antiTNF strategies that may offer more constrained or cell type specific control of TNF secretion are being sought. This requires identification of molecular mediators of TNF secretion, particularly those that can be targeted to block TNF release.We have characterized the secretory pathway for TNF in activated macrophages and identified key organelles and components of the trafficking machinery whose expression or function is up-regulated by LPS to support cytokine secretion (4-6). Among these are members of the SNARE family of membrane fusion proteins required at different points along this ...

show abstract

“…A common structural feature of golgins is a cytoplasmic coiled-coil domain (8) similar to that of the myosin family. These coiled-coil proteins localized in the Golgi complex were grouped as the golgin family, for example, giantin/macrogolgin/GCP372/GCP364 (6,7,9,10), golgin-245/p230 (11,12), p210 (13), golgin-160/GCP170 (14,15), golgin-97 (16), GM130/ golgin-95 (14,17), golgin-84 (18), and golgin-67 (19).…”

mentioning

confidence: 99%

Identification and Characterization of GCP16, a Novel Acylated Golgi Protein That Interacts with GCP170

Ohta

Misumi

Sohda

et al. 2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

GCP170, a member of the golgin family associated with the cytoplasmic face of the Golgi membrane, was found to have a Golgi localization signal at the NH 2 -terminal region (positions 137-237). Using this domain as bait in the yeast two-hybrid screening system, we identified a novel protein that interacted with GCP170. The 2.0-kilobase mRNA encoding a 137-amino acid protein of 16 kDa designated GCP16 was ubiquitously expressed. Immunofluorescence microscopy showed that GCP16 was co-localized with GCP170 and giantin in the Golgi region. Despite the absence of a hydrophobic domain sufficient for participating in membrane localization, GCP16 was found to be tightly associated with membranes like an integral membrane protein. 69 and Cys 72 , accounting for its tight association with the membrane. A mutant without potential acylation sites (C69A/C72A) was no longer localized to the Golgi, indicating that the acylation is prerequisite for the Golgi localization of GCP16. Although the mutant GCP16, even when overexpressed, had no effect on protein transport, overexpression of the wild type GCP16 caused an inhibitory effect on protein transport from the Golgi to the cell surface. Taken together, these results indicate that GCP16 is the acylated membrane protein, associated with GCP170, and possibly involved in vesicular transport from the Golgi to the cell surface. Labeling experiments with [ 3 H]palmitic acid and mutational analysis demonstrated that GCP16 was acylated at Cys

show abstract

Molecular cloning of a novel 97‐kd Golgi complex autoantigen associated with Sjögren's syndrome

Cited by 157 publications

References 49 publications

A Phosphorylated Cytoplasmic Autoantigen, GW182, Associates with a Unique Population of Human mRNAs within Novel Cytoplasmic Speckles

A Phosphorylated Cytoplasmic Autoantigen, GW182, Associates with a Unique Population of Human mRNAs within Novel Cytoplasmic Speckles

A trans -Golgi network golgin is required for the regulated secretion of TNF in activated macrophages in vivo

Identification and Characterization of GCP16, a Novel Acylated Golgi Protein That Interacts with GCP170

Contact Info

Product

Resources

About