| 1993 |
Giantin is an integral Golgi membrane protein with a large cytoplasmic domain (up to 350 kDa accessible to proteolysis from intact Golgi vesicles), a disulfide-linked lumenal domain, and a C-terminal transmembrane anchor; its topology and conservation suggest a role in forming intercisternal cross-bridges of the Golgi complex. |
Differential centrifugation, sucrose flotation, carbonate extraction, limited proteolysis of intact Golgi vesicles, non-reducing SDS-PAGE, double immunofluorescence with galactosyltransferase marker |
Molecular biology of the cell |
High |
7691276
|
| 1994 |
Giantin is a 376 kDa Golgi membrane protein with an extraordinarily high content of heptad repeats, consistent with coiled-coil structure similar to myosin family proteins, localized to the Golgi complex by immunoelectron microscopy. |
cDNA cloning, sequence analysis of heptad repeats, immunoelectron microscopy, immunofluorescence with brefeldin A treatment, subcellular fractionation |
Molecular and cellular biology |
High |
7511208
|
| 1998 |
Giantin is present on COPI vesicles and acts as a vesicle-side receptor for p115; the giantin–p115–GM130 tethering complex docks COPI vesicles to Golgi membranes, with giantin on the vesicle, GM130 on the Golgi membrane, and p115 bridging them. |
Immunoprecipitation of p115 binding partners from detergent Golgi extracts, antibody inhibition of vesicle docking in cell-free assay, GM130 peptide competition, immunodepletion |
The Journal of cell biology |
High |
9490716
|
| 2000 |
The N-terminal 15% of giantin is sufficient to bind p115 both in vitro and in vivo, and this fragment blocks cell-free Golgi reassembly, consistent with a long flexible tether linking COPI vesicles to cisternae. |
In vitro binding assay with recombinant N-terminal giantin fragments, co-immunoprecipitation in vivo, cell-free Golgi reassembly inhibition assay |
The Journal of biological chemistry |
High |
10644749
|
| 2000 |
Giantin is required for ER-to-Golgi transport: giantin p115-binding domain peptides and anti-giantin antibodies inhibit VSV-G protein transport to the mannosidase II-containing Golgi compartment at a step temporally after the GM130-requiring step, indicating giantin acts sequentially downstream of GM130 in ER-Golgi trafficking. |
VSV-G transport assay, inhibitory peptide injection, antibody microinjection, kinetic comparison with anti-p115 and anti-GM130 reagents |
The Journal of biological chemistry |
High |
11035033
|
| 2001 |
The Golgi localization signal of giantin resides in its C-terminal cytoplasmic domain; a novel peripheral Golgi protein GCP60 interacts with this C-terminal domain of giantin via its own C-terminal domain, and overexpression of the GCP60 C-terminal domain causes Golgi disassembly and blocks ER-to-Golgi transport. |
Yeast two-hybrid screening using giantin C-terminal cytoplasmic domain as bait, co-immunoprecipitation, immunofluorescence, immunoelectron microscopy, overexpression dominant-negative analysis |
The Journal of biological chemistry |
Medium |
11590181
|
| 2001 |
In vivo, reduction of p115 below detectable levels causes COPI-dependent Golgi fragmentation; however, reducing giantin below detectable levels or inhibiting p115 binding to GM130 has no detectable effect on Golgi structure or reassembly after cell division or brefeldin A washout, demonstrating that p115's essential role in Golgi structure is independent of giantin and GM130. |
Antibody microinjection targeting mapped binding sites, proteasome-mediated antigen depletion, immunofluorescence for Golgi structure, COPI inhibition co-treatment |
The Journal of cell biology |
Medium |
11591729
|
| 2007 |
Giantin interacts with the small GTPase Rab6A (in addition to previously known Rab1) both in vivo (co-immunoprecipitation) and in vitro (pulldown), suggesting that two distinct Rab GTPases can bind to the same golgin protein. |
Co-immunoprecipitation from cells, in vitro pulldown assay |
Experimental cell research |
Medium |
17475246
|
| 2011 |
A 10-bp insertion in exon 13 of Golgb1 causing a frameshift and premature stop at codon 1082 (truncating the C-terminal two-thirds including the Golgi-targeting region) abolishes giantin protein expression and causes osteochondrodysplasia, systemic edema, cleft palate, and lethal dwarfism in ocd/ocd rats, establishing giantin as essential for chondrogenesis. |
Fine linkage mapping, sequence analysis of mutant allele, in-gel Western blotting with C-terminal epitope antibody, histology, immunohistochemistry of growth plate |
Bone |
High |
21851869
|
| 2013 |
Giantin is required for ciliogenesis: siRNA-mediated depletion of giantin causes mis-localization of WDR34 (dynein-2 intermediate chain) and prevents primary cilia formation; partial depletion increases cilia length, consistent with giantin controlling ciliogenesis through regulation of dynein-2 localization rather than through the Rab11-Rabin8-Rab8 ciliary membrane pathway. |
siRNA knockdown, immunofluorescence for cilia markers and WDR34, cilia length measurement, epistasis with Rab11/Rabin8/Rab8 pathway components |
Journal of cell science |
Medium |
24046448
|
| 2013 |
Giantin mediates the spatial organization of the Golgi ribbon: siRNA depletion of giantin results in more dispersed Golgi mini-stacks after nocodazole treatment without changing cisternal length; exogenous expression of mammalian giantin in Drosophila S2 cells (which lack a giantin homolog and have dispersed stacks) induces clustering of Golgi stacks, demonstrating that giantin is sufficient and necessary for Golgi ribbon organization. |
siRNA knockdown, nocodazole treatment, immunofluorescence, exogenous expression of giantin cDNA in Drosophila S2 cells, glycosylation assays |
PloS one |
Medium |
23555793
|
| 2016 |
Golgb1 loss-of-function mutation in mice (confirmed by CRISPR/Cas9-generated alleles) causes cleft palate with intrinsic defects in palatal shelf elevation: mutant palatal mesenchyme shows increased cell density, reduced hyaluronan accumulation, and impaired protein glycosylation, demonstrating a specific role for giantin in protein glycosylation and tissue morphogenesis. |
ENU mutagenesis screen, genetic linkage mapping, whole-exome sequencing, CRISPR/Cas9 genome editing, maxillary explant culture, immunostaining for hyaluronan and glycosylation markers |
Development (Cambridge, England) |
High |
27226319
|
| 2017 |
Giantin knockout leads to near-complete loss of GALNT3 function and differential expression of 22 Golgi-resident glycosyltransferases (but not glycan-processing enzymes or ER glycosylation machinery); giantin-knockout zebrafish phenocopy hyperphosphatemic familial tumoral calcinosis (caused by GALNT3 mutations), establishing a feedback loop between Golgi structure and glycosyltransferase expression. |
Giantin knockout in mammalian cells and zebrafish (CRISPR/Cas9), RNA-seq, phenotypic characterization (hyperostosis, ectopic calcium deposits), comparison to GALNT3 disease phenotype |
Journal of cell science |
High |
29093022
|
| 2017 |
In androgen-independent prostate cancer cells with defective giantin, Golgi targeting of glycosyltransferases and α-mannosidase IA shifts from giantin to GM130-GRASP65; this results in acquisition of high mannose N-glycans on trans-Golgi enzymes and cell surface glycoproteins, absent in cells with functional giantin. |
Confocal microscopy after knockdown of GM130 or giantin, in situ proximity ligation assay, MALDI-TOF mass spectrometry of N-glycans from immunoprecipitated enzyme |
Biochimica et biophysica acta. General subjects |
Medium |
28782625
|
| 2017 |
Giantin loss-of-function in zebrafish causes elongated cilia in the neural tube and accumulation of material at the ciliary tip, consistent with defective retrograde intraflagellar transport, corroborating a role for giantin in ciliogenesis through retrograde dynein-2-mediated transport. |
Morpholino knockdown, CRISPR/Cas9 knockout in zebrafish, scanning electron microscopy of cilia, cilia number and length measurements |
Biology open |
Medium |
28546340
|
| 2018 |
Giantin is required for coordinated production of aggrecan, link protein and type XI collagen in chondrocytes at the post-transcriptional/secretory level: ocd/ocd rat chondrocytes lacking giantin show reduced aggrecan and link protein and increased type XI collagen protein levels despite normal mRNA levels, indicating giantin is required for proper secretion/trafficking of these ECM components. |
Immunostaining of embryonic femur cartilage from ocd/ocd and normal rats, semi-quantitative RT-PCR for ECM mRNAs, Safranin O/azan staining of epiphyseal cartilage, isolated chondrocyte cultures |
Biochemical and biophysical research communications |
Medium |
29577904
|
| 2018 |
Giantin exists as a dimer linked by a disulfide bond in its luminal domain; ethanol-induced Golgi disorganization is associated with giantin de-dimerization, and post-ethanol Golgi recovery requires giantin re-dimerization as well as Rab6a GTPase; knockdown of giantin, Rab6a, or non-muscle myosin IIB impairs post-ethanol Golgi recovery. |
Proximity ligation assay for giantin dimerization state, siRNA knockdown, immunofluorescence for Golgi structure, ethanol treatment/withdrawal model in VA-13 cells and rat hepatocytes |
Biomolecules |
Medium |
30453527
|
| 2019 |
Giantin knockdown reduces fenestrae within Golgi cisternae and increases diffusion rate of Golgi membrane proteins, indicating increased connectivity among cisternae and stacks; this suggests giantin inhibits rather than promotes tethering/fusion of nearby Golgi cisternae, contrary to the classical cis-golgin tether model. |
siRNA knockdown, electron tomography and 3D modeling of Golgi, FRAP measurement of Golgi membrane protein diffusion, glycosylation assays |
Frontiers in cell and developmental biology |
Medium |
31544102
|
| 2019 |
Post-BFA stress Golgi reassembly is governed by giantin re-dimerization via its luminal disulfide bond assisted by Rab6a GTPase; giantin-sensitive Golgi resident enzymes are recruited to nascent Golgi membranes after complete recovery via direct interaction of their cytoplasmic tails with the N-terminus of giantin, whereas GM130-GRASP65-dependent enzymes arrive earlier. |
Brefeldin A treatment/washout model, proximity ligation assay, atomic force microscopy of giantin conformational state, 3D SIM super-resolution microscopy |
Cells |
Medium |
31847122
|
| 2021 |
Giantin is required for intracellular N-terminal propeptide processing of type I procollagen: in human giantin-knockout cells expressing GFP-tagged procollagen, procollagen trafficking is independent of giantin but N-propeptide cleavage of pro-α1(I) is defective; giantin-mutant zebrafish accumulate spontaneous fractures and show defective mineralization of newly deposited collagen. |
Human giantin-knockout cell line, GFP-tagged procollagen trafficking assay, immunoblot for N-propeptide processing, zebrafish giantin mutant fracture induction and mineralization assay, procollagen reporter expression |
The Journal of cell biology |
High |
33944912
|
| 2024 |
Giantin forms protein complexes with Gal3-O-sulfotransferases (Gal3STs) and is required for their proper Golgi localization; loss or mislocalization of giantin (as occurs in salivary glands of Sjögren's disease patients) alters Gal3ST localization, reduces Gal3ST activity, and decreases sulfation of MUC5B mucin. |
Co-immunoprecipitation of giantin-Gal3ST complexes, giantin knockout and knockdown cell lines, immunofluorescence localization, MUC5B sulfation analysis from patient salivary gland biopsies |
JCI insight |
Medium |
39388276
|
| 2026 |
GOLGB1 knockdown activates the MAPK pathway (elevated p-ERK and p-P38), promotes nucleus pulposus cell degeneration and apoptosis, and inhibits proliferation; MAPK inhibitor SCH772984 rescues the degeneration phenotype, positioning GOLGB1 upstream of MAPK signaling in intervertebral disc homeostasis. |
Plasmid transfection (knockdown), Western blotting for p-ERK/p-P38, EdU proliferation assay, TUNEL/flow cytometry apoptosis assay, CCK8, mouse IDD model with histology/IHC, MAPK inhibitor rescue |
Scientific reports |
Medium |
41547890
|