An NSF function distinct from ATPase-dependent SNARE disassembly is essential for Golgi membrane fusion

10.1038/14025 ◽  
1999 ◽  
Vol 1 (6) ◽  
pp. 335-340 ◽  
Author(s):  
Joyce M. M. Müller ◽  
Catherine Rabouille ◽  
Richard Newman ◽  
James Shorter ◽  
Paul Freemont ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Golgi Membrane

scholarly journals DjA1 maintains Golgi integrity via interaction with GRASP65

2019 ◽  
Vol 30 (4) ◽  
pp. 478-490 ◽  
Author(s):  
Jie Li ◽  
Danming Tang ◽  
Stephen C. Ireland ◽  
Yanzhuang Wang
Keyword(s):  
Heat Shock ◽  
Membrane Fusion ◽  
Structure Formation ◽  
Mammalian Cells ◽  
Binding Protein ◽  
Golgi Membrane ◽  
Biochemical Methods ◽  
Golgi Fragmentation ◽  
Golgi Structure

In mammalian cells, the Golgi reassembly stacking protein of 65 kDa (GRASP65) has been implicated in both Golgi stacking and ribbon linking by forming trans-oligomers. To better understand its function and regulation, we used biochemical methods to identify the DnaJ homolog subfamily A member 1 (DjA1) as a novel GRASP65-binding protein. In cells, depletion of DjA1 resulted in Golgi fragmentation, short and improperly aligned cisternae, and delayed Golgi reassembly after nocodazole washout. In vitro, immunodepletion of DjA1 from interphase cytosol reduced its activity to enhance GRASP65 oligomerization and Golgi membrane fusion, while adding purified DjA1 enhanced GRASP65 oligomerization. DjA1 is a cochaperone of Heat shock cognate 71-kDa protein (Hsc70), but the activity of DjA1 in Golgi structure formation is independent of its cochaperone activity or Hsc70, rather, through DjA1-GRASP65 interaction to promote GRASP65 oligomerization. Thus, DjA1 interacts with GRASP65 to enhance Golgi structure formation through the promotion of GRASP65 trans-oligomerization.

scholarly journals Sequential SNARE disassembly and GATE-16–GOS-28 complex assembly mediated by distinct NSF activities drives Golgi membrane fusion

2002 ◽  
Vol 157 (7) ◽  
pp. 1161-1173 ◽  
Author(s):  
Joyce M.M. Müller ◽  
James Shorter ◽  
Richard Newman ◽  
Katrin Deinhardt ◽  
Yuval Sagiv ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Atp Hydrolysis ◽  
Golgi Membrane ◽  
Complex Assembly ◽  
Evolutionarily Conserved ◽  
Golgi Fragmentation ◽  
Fusion Analysis

Characterization of mammalian NSF (G274E) and Drosophila NSF (comatose) mutants revealed an evolutionarily conserved NSF activity distinct from ATPase-dependent SNARE disassembly that was essential for Golgi membrane fusion. Analysis of mammalian NSF function during cell-free assembly of Golgi cisternae from mitotic Golgi fragments revealed that NSF disassembles Golgi SNAREs during mitotic Golgi fragmentation. A subsequent ATPase-independent NSF activity restricted to the reassembly phase is essential for membrane fusion. NSF/α-SNAP catalyze the binding of GATE-16 to GOS-28, a Golgi v-SNARE, in a manner that requires ATP but not ATP hydrolysis. GATE-16 is essential for NSF-driven Golgi reassembly and precludes GOS-28 from binding to its cognate t-SNARE, syntaxin-5. We suggest that this occurs at the inception of Golgi reassembly to protect the v-SNARE and regulate SNARE function.

Mitotic phosphorylation of VCIP135 blocks p97ATPase-mediated Golgi membrane fusion

2013 ◽  
Vol 433 (2) ◽  
pp. 237-242 ◽  
Author(s):  
Go Totsukawa ◽  
Ayaka Matsuo ◽  
Ayano Kubota ◽  
Yuya Taguchi ◽  
Hisao Kondo
Keyword(s):  
Membrane Fusion ◽  
Golgi Membrane ◽  
Mitotic Phosphorylation

scholarly journals Cell cycle regulation of VCIP135 deubiquitinase activity and function in p97/p47-mediated Golgi reassembly

2015 ◽  
Vol 26 (12) ◽  
pp. 2242-2251 ◽  
Author(s):  
Xiaoyan Zhang ◽  
Yanzhuang Wang
Keyword(s):  
Cell Cycle ◽  
Membrane Fusion ◽  
Mammalian Cells ◽  
Membrane Dynamics ◽  
Deubiquitinating Enzyme ◽  
Golgi Membrane ◽  
Interacting Protein ◽  
Daughter Cells ◽  
And Function ◽  
Cycle Regulation

In mammalian cells, the inheritance of the Golgi apparatus into the daughter cells during each cycle of cell division is mediated by a disassembly and reassembly process, and this process is precisely controlled by phosphorylation and ubiquitination. VCIP135 (valosin-containing protein p97/p47 complex–interacting protein, p135), a deubiquitinating enzyme required for p97/p47-mediated postmitotic Golgi membrane fusion, is phosphorylated at multiple sites during mitosis. However, whether phosphorylation directly regulates VCIP135 deubiquitinase activity and Golgi membrane fusion in the cell cycle remains unknown. We show that, in early mitosis, phosphorylation of VCIP135 by Cdk1 at a single residue, S130, is sufficient to inactivate the enzyme and inhibit p97/p47-mediated Golgi membrane fusion. At the end of mitosis, VCIP135 S130 is dephosphorylated, which is accompanied by the recovery of its deubiquitinase activity and Golgi reassembly. Our results demonstrate that phosphorylation and ubiquitination are coordinated via VCIP135 to control Golgi membrane dynamics in the cell cycle.

scholarly journals Acute inactivation of theAspergillus nidulans Golgi membrane fusion machinery: correlation of apical extension arrest and tip swelling with cisternal disorganization

2013 ◽  
Vol 89 (2) ◽  
pp. 228-248 ◽  
Author(s):  
Mario Pinar ◽  
Areti Pantazopoulou ◽  
Herbert N. Arst ◽  
Miguel A. Peñalva
Keyword(s):  
Membrane Fusion ◽  
Golgi Membrane

scholarly journals Mutations in Membrin/GOSR2 reveal stringent secretory pathway demands of dendritic growth and synaptic integrity

2017 ◽  
Author(s):  
Roman Praschberger ◽  
Simon A. Lowe ◽  
Nancy T. Malintan ◽  
Henry Houlden ◽  
Dimitri M. Kullmann ◽  
...  
Keyword(s):  
Nervous System ◽  
Membrane Fusion ◽  
Dendritic Growth ◽  
Secretory Pathway ◽  
Synaptic Function ◽  
Golgi Membrane ◽  
Snare Protein ◽  
Progressive Myoclonus Epilepsy ◽  
Myoclonus Epilepsy ◽  
Axonal Trafficking

AbstractMutations in the Golgi SNARE protein Membrin (encoded by the GOSR2 gene) cause progressive myoclonus epilepsy (PME). Membrin is a ubiquitously important protein mediating ER-to-Golgi membrane fusion, and hence it is unclear how these mutations result in a disorder restricted to the nervous system. Here we use a multi-layered strategy to elucidate the consequences of Membrin mutations from protein to neuron. We show that the pathogenic mutations cause partial reductions in SNARE-mediated membrane fusion. Importantly, these alterations were sufficient to profoundly impair dendritic growth in novel Drosophila models of GOSR2-PME. We also observed axonal trafficking abnormalities in this model, as well as synaptic malformations, trans-synaptic instability and hyperactive synaptic transmission. Our study highlights how dendritic growth is vulnerable even to subtle secretory pathway deficits, uncovers a previously uncharacterized role for Membrin in synaptic function, and provides a comprehensive explanatory basis for genotype-phenotype relationships in GOSR2-PME.

scholarly journals Phosphorylation regulates VCIP135 function in Golgi membrane fusion during the cell cycle

2013 ◽  
Vol 127 (1) ◽  
pp. 172-181 ◽  
Author(s):  
X. Zhang ◽  
H. Zhang ◽  
Y. Wang
Keyword(s):  
Cell Cycle ◽  
Membrane Fusion ◽  
Golgi Membrane

scholarly journals Membrane fusion and glycosylation in the rat hepatic Golgi apparatus.

1982 ◽  
Vol 92 (1) ◽  
pp. 147-154 ◽  
Author(s):  
J Paiement ◽  
R A Rachubinski ◽  
N M Ng Ying Kin ◽  
R A Sikstrom ◽  
J J Bergeron
Keyword(s):  
Electron Microscope ◽  
Membrane Fusion ◽  
Complete Medium ◽  
Transferase Activity ◽  
Golgi Membrane ◽  
Assay Medium ◽  
Galactosyl Transferase ◽  
Cacodylate Buffer ◽  
Triton X ◽  
Silver Grains

When purified Golgi fractions were incubated with UDP-[3H]galactose in the absence of Triton-X-100, radioactivity was incorporated into an endogenous lipid and several peptide acceptors. Electron microscope analysis of Golgi fractions incubated in the endogenous galactosyl transferase assay medium revealed extensive fusion of Golgi saccules. Systematic removal of constituents in the galactosyl transferase assay medium showed enhanced (minus beta-mercaptoethanol) or reduced (minus ATP, minus sodium cacodylate buffer or minus MnCl2) fusion of Golgi membranes compared to the complete medium, Stereologic analysis revealed a correlation between membrane fusion and galactosyl transferase activity (r = 0.99, P less than 0.001). Electron microscope radioautography was carried out after incubation of Golgi fractions with UDP-[3H]galactose. Silver grains were not observed over trans elements of Golgi but were revealed mainly over large fused saccules with the number of silver grains being proportionate to membrane fusion (r = 0.92, P less than 0.001). Bilayer destabilization at points of Golgi membrane fusion may act to translocate galactose across the Golgi membrane and thereby provide a fusion regulated substrate for terminal glycosylation.

scholarly journals Mena–GRASP65 interaction couples actin polymerization to Golgi ribbon linking

2016 ◽  
Vol 27 (1) ◽  
pp. 137-152 ◽  
Author(s):  
Danming Tang ◽  
Xiaoyan Zhang ◽  
Shijiao Huang ◽  
Hebao Yuan ◽  
Jie Li ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Mammalian Cells ◽  
Actin Polymerization ◽  
Binding Protein ◽  
Elongation Factor ◽  
Golgi Membrane ◽  
Golgi Membranes ◽  
Golgi Fragmentation ◽  
Golgi Ribbon

In mammalian cells, the Golgi reassembly stacking protein 65 (GRASP65) has been implicated in both Golgi stacking and ribbon linking by forming trans-oligomers through the N-terminal GRASP domain. Because the GRASP domain is globular and relatively small, but the gaps between stacks are large and heterogeneous, it remains puzzling how GRASP65 physically links Golgi stacks into a ribbon. To explore the possibility that other proteins may help GRASP65 in ribbon linking, we used biochemical methods and identified the actin elongation factor Mena as a novel GRASP65-binding protein. Mena is recruited onto the Golgi membranes through interaction with GRASP65. Depleting Mena or disrupting actin polymerization resulted in Golgi fragmentation. In cells, Mena and actin were required for Golgi ribbon formation after nocodazole washout; in vitro, Mena and microfilaments enhanced GRASP65 oligomerization and Golgi membrane fusion. Thus Mena interacts with GRASP65 to promote local actin polymerization, which facilitates Golgi ribbon linking.

Sign in / Sign up

Export Citation Format

Share Document