Rapid degradation of GRASP55 and GRASP65 reveals their immediate impact on the Golgi structure.
Journal
The Journal of cell biology
ISSN: 1540-8140
Titre abrégé: J Cell Biol
Pays: United States
ID NLM: 0375356
Informations de publication
Date de publication:
04 01 2021
04 01 2021
Historique:
received:
09
07
2020
revised:
20
10
2020
accepted:
28
10
2020
entrez:
10
12
2020
pubmed:
11
12
2020
medline:
19
5
2021
Statut:
ppublish
Résumé
GRASP55 and GRASP65 have been implicated in stacking of Golgi cisternae and lateral linking of stacks within the Golgi ribbon. However, RNAi or gene knockout approaches to dissect their respective roles have often resulted in conflicting conclusions. Here, we gene-edited GRASP55 and/or GRASP65 with a degron tag in human fibroblasts, allowing for induced rapid degradation by the proteasome. We show that acute depletion of either GRASP55 or GRASP65 does not affect the Golgi ribbon, while chronic degradation of GRASP55 disrupts lateral connectivity of the ribbon. Acute double depletion of both GRASPs coincides with the loss of the vesicle tethering proteins GM130, p115, and Golgin-45 from the Golgi and compromises ribbon linking. Furthermore, GRASP55 and/or GRASP65 is not required for maintaining stacks or de novo assembly of stacked cisternae at the end of mitosis. These results demonstrate that both GRASPs are dispensable for Golgi stacking but are involved in maintaining the integrity of the Golgi ribbon together with GM130 and Golgin-45.
Identifiants
pubmed: 33301566
pii: 211583
doi: 10.1083/jcb.202007052
pmc: PMC7735681
pii:
doi:
Substances chimiques
GORASP1 protein, human
0
GORASP2 protein, human
0
Golgi Matrix Proteins
0
Indoleacetic Acids
0
Brefeldin A
20350-15-6
Nocodazole
SH1WY3R615
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM096070
Pays : United States
Organisme : NIH HHS
ID : S10 OD021685
Pays : United States
Informations de copyright
© 2020 Zhang and Seemann.
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