Reconstitution of Phospholipase A2-Dependent Golgi Membrane Tubules

Cluett, Edward B.; Figueiredo, Paul de; Bechler, Marie E.; Thorsen, Kevin; Brown, William J.

doi:10.1007/978-1-4939-6463-5_7

Cited by 2 publications

(2 citation statements)

References 43 publications

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“…To validate these screens, we implemented the proof-of-principle experiment, using cell-free reconstitution of Golgi membranes [97]. First, we isolated Golgi membranes from non-treated and BFA-treated HeLa cells according to the protocol established previously [29].…”

Section: The Mutuality Of Rab6a and Giantinmentioning

confidence: 99%

Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

Frisbie

Lushnikov

Krasnoslobodtsev

et al. 2019

Cells

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Background: The Golgi apparatus undergoes disorganization in response to stress, but it is able to restore compact and perinuclear structure under recovery. This self-organization mechanism is significant for cellular homeostasis, but remains mostly elusive, as does the role of giantin, the largest Golgi matrix dimeric protein. Methods: In HeLa and different prostate cancer cells, we used the model of cellular stress induced by Brefeldin A (BFA). The conformational structure of giantin was assessed by proximity ligation assay and atomic force microscopy. The post-BFA distribution of Golgi resident enzymes was examined by 3D SIM high-resolution microscopy. Results: We detected that giantin is rather flexible than an extended coiled-coil dimer and BFA-induced Golgi disassembly was associated with giantin monomerization. A fusion of the nascent Golgi membranes after BFA washout is forced by giantin re-dimerization via disulfide bond in its luminal domain and assisted by Rab6a GTPase. GM130-GRASP65-dependent enzymes are able to reach the nascent Golgi membranes, while giantin-sensitive enzymes appeared at the Golgi after its complete recovery via direct interaction of their cytoplasmic tail with N-terminus of giantin. Conclusion: Post-stress recovery of Golgi is conducted by giantin dimer and Golgi proteins refill membranes according to their docking affiliation rather than their intra-Golgi location.

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Section: The Mutuality Of Rab6a and Giantinmentioning

confidence: 99%

Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

Frisbie

Lushnikov

Krasnoslobodtsev

et al. 2019

Cells

View full text Add to dashboard Cite

show abstract

“…To validate these screens, we implemented the proof-of-principle experiment, using cell-free reconstitution of Golgi membrane tubule formation (Cluett, de Figueiredo et al, 2016). First, we isolated Golgi membranes from non-treated and BFA-treated HeLa cells according to the protocol established previously (Petrosyan & Cheng, 2013).…”

Section: Resultsmentioning

confidence: 99%

Golgi Renaissance: the pivotal role of the largest Golgi protein giantin

Petrosyan

Manca

Casey

et al. 2018

Preprint

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Golgi undergoes disorganization in response to the drugs or alcohol, but it is able to restore compact structure under recovery. This self-organization mechanism remains mostly elusive, as does the role of giantin, the largest Golgi matrix dimeric protein. Here, we found that in cells treated with Brefeldin A (BFA) or ethanol (EtOH), Golgi disassembly is associated with giantin de-dimerization, which was restored to the dimer form after BFA or EtOH washout. Cells lacking giantin are disabled for the restoration of the classical ribbon Golgi, and they demonstrate altered trafficking of proteins to the cell surface. The fusion of the nascent Golgi membranes is mediated by the cross-membrane interaction of Rab6a GTPase and giantin. Giantin is involved in the formation of long intercisternal connections, which in giantin-depleted cells was replaced by the short bridges that formed via oligomerization of GRASP65. This phenomenon occurs in advanced prostate cancer cells, in which a fragmented Golgi phenotype is maintained by the dimerization of GRASP65. Thus, we provide a model of Golgi Renaissance, which is impaired in aggressive prostate cancer.

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Reconstitution of Phospholipase A2-Dependent Golgi Membrane Tubules

Cited by 2 publications

References 43 publications

Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

Post-ER Stress Biogenesis of Golgi Is Governed by Giantin

Golgi Renaissance: the pivotal role of the largest Golgi protein giantin

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