Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis

• DOI: 10.1073/pnas.0306373101
• Publication Date: 2004-02-03T17:47:24Z
• Authors: Manojkumar A. Put...; Adam D. Linstedt
• Keywords: Transport protein

Functional characterization of protein interactions in mammalian systems has been hindered by the inability to perform complementation analyses vivo. Here, we use functional replacement vesicle docking p115 separate its essential from nonessential interactions. is required for biogenesis Golgi apparatus, but it unclear whether mechanism action requires golgin and/or SNARE Short interfering RNA-mediated knockdown induced extensive fragmentation and impaired secretory traffic. Reassembly a structurally functionally normal occurred on expression homologue not recognized short RNA. Strikingly, versions lacking phosphorylation site golgin-binding domains also restored apparatus cells endogenous p115. In contrast, SNARE-interacting domain was biogenesis. This suggests that acts directly, rather than via tether, catalyze trans-SNARE complex formation preceding membrane fusion.