Handling Editor: David Glover
ABSTRACT
Golgi stacks are the basic structural units of the Golgi. Golgi stacks are separated from each other and scattered in the cytoplasm of Drosophila cells. Here, we report that the ARF-GEF inhibitor Brefeldin A (BFA) induces the formation of BFA bodies, which are aggregates of Golgi stacks, trans-Golgi networks and recycling endosomes. Recycling endosomes are located in the centers of BFA bodies, while Golgi stacks surround them on their trans sides. Live imaging of S2 cells revealed that Golgi stacks repeatedly merged and separated on their trans sides, and BFA caused successive merger by inhibiting separation, forming BFA bodies. S2 cells carrying genome-edited BFA-resistant mutant Sec71M717L did not form BFA bodies at high concentrations of BFA; S2 cells carrying genome-edited BFA-hypersensitive mutant Sec71F713Y produced BFA bodies at low concentrations of BFA. These results indicate that Sec71 is the sole BFA target for BFA body formation and controls Golgi stack separation. Finally, we showed that impairment of Sec71 in fly photoreceptors induces BFA body formation, with accumulation of both apical and basolateral cargoes, resulting in inhibition of polarized transport.
Footnotes
Competing interests
The authors declare no competing or financial interests.
Author contributions
Conceptualization: T.S., A.K.S.; Methodology: S.F., K.K., T.S., A.K.S.; Formal analysis: S.F.; Investigation: S.F., K.K., T.T., R.I., A.T., T.S.; Data curation: S.F., T.S., A.K.S.; Writing - original draft: T.S., A.K.S.; Writing - review & editing: K.K., A.N.; Supervision: A.N., T.S., A.K.S.; Project administration: T.S., A.K.S.; Funding acquisition: K.K., A.N., T.S., A.K.S.
Funding
This work was supported by Precursory Research for Embryonic Science and Technology grant 25-J-J4215, Japan Society for the Promotion of Science, KAKENHI grant 15K07050, Sumitomo Foundation for Basic Science Research Projects, Astellas Foundation for Research on Metabolic Disorders, and Female Researcher Joint Research Grant from Hiroshima University to A.S., KAKENHI grant 19K06566 to T.S., and KAKENHI grants 25221103, 17H06420, and 18H05275 to K.K. and A.N.
Supplementary information
Supplementary information available online at https://jcs.biologists.org/lookup/doi/10.1242/jcs.245571.supplemental
- Received May 18, 2020.
- Accepted November 18, 2020.
- © 2020. Published by The Company of Biologists Ltd
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