Microtubule-independent secretion requires functional maturation of Golgi elements

The Golgi apparatus is responsible for processing and sorting of secretory cargos. Microtubules are known to accelerate the transport of proteins from the endoplasmic reticulum to the Golgi apparatus and from the Golgi to the plasma membrane. However, whether post-Golgi transport strictly requires microtubules is still unclear. Using the retention using selective hooks (RUSH) system to synchronize the trafficking of cargos, we show that anterograde transport of tumor necrosis factor (TNF) is strongly reduced without microtubules. We show that two populations of Golgi elements co-exist in these cells. A centrally located and giantin-positive Golgi complex sustains trafficking while newly formed peripheral Golgi mini-stacks accumulate cargos in cells without microtubules. Using a genome-edited GFP-giantin cell line, we observe that the trafficking-competent Golgi population corresponds to the pre-existing one that was present before removal of microtubules. All Golgi elements support trafficking after long-term microtubules depletion or after relocation of Golgi proteins in the endoplasmic reticulum using Brefeldin A. Our results demonstrate that functional maturation of Golgi elements is needed to ensure post-Golgi trafficking and that microtubule-driven post-Golgi transport is not strictly required.