De novo Golgi biogenesis requires orchestrated transactivation of Golgi genes

Golgi biogenesis is a crucial cellular process, yet its study has been hindered by the lack of easy methods to induce massive production of new Golgi units. In this study, we optimized an enzyme inactivation approach to stimulate the formation of fully functional Golgi stacks. Massive Golgi biogenesis was achieved through DAB-mediated inactivation of preexisting Golgi structures in cells stably expressing the chimeric Golgi enzyme Mannosidase II fused with horse radish peroxidase (HRP). Within 10 to 18 hours post-Golgi inactivation, substantial number of cells regenerated new Golgi stacks capable of transporting various cargo proteins to their appropriate destinations. This functional and structural recovery of the Golgi in ManII-HRP cells offers an efficient system to study Golgi biogenesis and associated cellular responses. We further explored transcriptomic changes in ManII-HRP cells, discovering that the extensive biogenesis of Golgi membranes induces the transcriptional activation of numerous Golgi-associated genes. These genes encode proteins involved in diverse functions such as glycosylation, membrane tethering, and ion/sugar transport. The timing of this gene transactivation correlates with the restoration of Golgi cisternae architecture and trafficking capabilities, suggesting that this transcriptional response plays an integral role in the biogenesis of new Golgi units.