Reconstitution of the cell cycle-regulated Golgi disassembly and reassembly in a cell-free system

Tang, Danming; Xiang, Ye; Wang, Yanzhuang

doi:10.1038/nprot.2010.38

Cited by 50 publications

(84 citation statements)

References 49 publications

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“…For analysis, Golgi stacks were identified using morphological criteria and quantified using standard stereological techniques (59,60). Golgi images were captured at ×11,000 magnification.…”

Section: Discussionmentioning

confidence: 99%

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Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Tan

Banerjee

Guo

et al. 2016

Journal of Clinical Investigation

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Section: Discussionmentioning

confidence: 99%

“…Electron microscopy was performed as previously described (59,60). Sections of 60 nm were mounted on Formvar-coated nickel grids and double contrasted with 2% uranyl acetate for 5 minutes and 3% lead citrate for 5 minutes.…”

Section: Discussionmentioning

confidence: 99%

Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Tan

Banerjee

Guo

et al. 2016

Journal of Clinical Investigation

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“…To demonstrate functional importance beyond ATPase activities, we showed that a disease mutant is hyperactive and less responsive to activation by p37 in postmitotic Golgi membrane fusion in a well-characterized in vitro reconstitution assay (36,47). Specifically, the R155H disease mutant is 2.5 times more active than WT in this p97-mediated postmitotic Golgi reassembly assay (SI Appendix, Fig.…”

Section: δ69-92 P47 Can Bind To P97 Disease Mutants But Cannot Activatementioning

confidence: 99%

Altered cofactor regulation with disease-associated p97/VCP mutations

Zhang

Gui

Zhang

et al. 2015

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

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147

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Dominant mutations in p97/VCP (valosin-containing protein) cause a rare multisystem degenerative disease with varied phenotypes that include inclusion body myopathy, Paget's disease of bone, frontotemporal dementia, and amyotrophic lateral sclerosis. p97 disease mutants have altered N-domain conformations, elevated ATPase activity, and altered cofactor association. We have now discovered a previously unidentified disease-relevant functional property of p97 by identifying how the cofactors p37 and p47 regulate p97 ATPase activity. We define p37 as, to our knowledge, the first known p97-activating cofactor, which enhances the catalytic efficiency (k cat /K m ) of p97 by 11-fold. Whereas both p37 and p47 decrease the K m of ATP in p97, p37 increases the k cat of p97. In contrast, regulation by p47 is biphasic, with decreased k cat at low levels but increased k cat at higher levels. By deleting a region of p47 that lacks homology to p37 (amino acids 69-92), we changed p47 from an inhibitory cofactor to an activating cofactor, similar to p37. Our data suggest that cofactors regulate p97 ATPase activity by binding to the N domain. Induced conformation changes affect ADP/ATP binding at the D1 domain, which in turn controls ATPase cycling. Most importantly, we found that the D2 domain of disease mutants failed to be activated by p37 or p47. Our results show that cofactors play a critical role in controlling p97 ATPase activity, and suggest that lack of cofactorregulated communication may contribute to p97-associated disease pathogenesis.AAA ATPase | p97/VCP | MSP1 | p47 | steady-state kinetics

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“…The aim of this study was to identify the dynamic behaviors of the Golgi apparatus by physical modeling and to determine whether the process is mediated by self-organization. In particular, we focused on the Golgi reassembly process for the following reasons: (i) The Golgi reassembly process involves building of the whole Golgi structure de novo, and (ii) an in vitro assay of Golgi reassembly from Golgi-derived vesicles has been developed successfully (18,19). The second reason implies that…”

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confidence: 99%

Golgi apparatus self-organizes into the characteristic shape via postmitotic reassembly dynamics

Tachikawa

Mochizuki

2017

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

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The Golgi apparatus is a membrane-bounded organelle with the characteristic shape of a series of stacked flat cisternae. During mitosis in mammalian cells, the Golgi apparatus is once fragmented into small vesicles and then reassembled to form the characteristic shape again in each daughter cell. The mechanism and details of the reassembly process remain elusive. Here, by the physical simulation of a coarse-grained membrane model, we reconstructed the threedimensional morphological dynamics of the Golgi reassembly process. Considering the stability of the interphase Golgi shape, we introduce two hypothetical mechanisms-the Golgi rim stabilizer protein and curvature-dependent restriction on membrane fusioninto the general biomembrane model. We show that the characteristic Golgi shape is spontaneously organized from the assembly of vesicles by proper tuning of the two additional mechanisms, i.e., the Golgi reassembly process is modeled as self-organization. We also demonstrate that the fine Golgi shape forms via a balance of three reaction speeds: vesicle aggregation, membrane fusion, and shape relaxation. Moreover, the membrane fusion activity decreases thickness and the number of stacked cisternae of the emerging shapes.Golgi apparatus | self-organization | physical biology modeling | computer simulation

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Reconstitution of the cell cycle-regulated Golgi disassembly and reassembly in a cell-free system

Cited by 50 publications

References 49 publications

Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Altered cofactor regulation with disease-associated p97/VCP mutations

Golgi apparatus self-organizes into the characteristic shape via postmitotic reassembly dynamics

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