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.
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4 January 2021
Article|
December 10 2020
Rapid degradation of GRASP55 and GRASP65 reveals their immediate impact on the Golgi structure
Yijun Zhang
,
Yijun Zhang
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX
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Joachim Seemann
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX
Correspondence to Joachim Seemann: joachim.seemann@utsouthwestern.edu
Search for other works by this author on:
Yijun Zhang
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX
Joachim Seemann
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX
Correspondence to Joachim Seemann: joachim.seemann@utsouthwestern.edu
Received:
July 09 2020
Revision Received:
October 20 2020
Accepted:
October 28 2020
Online Issn: 1540-8140
Print Issn: 0021-9525
Funding:
National Institutes of Health
(GM096070)
Welch Foundation
(I-1910)
© 2020 Zhang and Seemann
2020
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Cell Biol (2021) 220 (1): e202007052.
Article history
Received:
July 09 2020
Revision Received:
October 20 2020
Accepted:
October 28 2020
Citation
Yijun Zhang, Joachim Seemann; Rapid degradation of GRASP55 and GRASP65 reveals their immediate impact on the Golgi structure. J Cell Biol 4 January 2021; 220 (1): e202007052. doi: https://doi.org/10.1083/jcb.202007052
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