de Figueiredo P, Polizotto R S, Drecktrah D, Brown W J
Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, New York 14853, USA.
Mol Biol Cell. 1999 Jun;10(6):1763-82. doi: 10.1091/mbc.10.6.1763.
Although membrane tubules can be found extending from, and associated with, the Golgi complex of eukaryotic cells, their physiological function has remained unclear. To gain insight into the biological significance of membrane tubules, we have developed methods for selectively preventing their formation. We show here that a broad range of phospholipase A2 (PLA2) antagonists not only arrest membrane tubule-mediated events that occur late in the assembly of the Golgi complex but also perturb its normal steady-state tubulovesicular architecture by inducing a reversible fragmentation into separate "mini-stacks." In addition, we show that these same compounds prevent the formation of membrane tubules from Golgi stacks in an in vitro reconstitution system. This in vitro assay was further used to demonstrate that the relevant PLA2 activity originates from the cytoplasm. Taken together, these results demonstrate that Golgi membrane tubules, sensitive to potent and selective PLA2 antagonists, mediate both late events in the reassembly of the Golgi complex and the dynamic maintenance of its steady-state architecture. In addition, they implicate a role for cytoplasmic PLA2 enzymes in mediating these membrane trafficking events.
尽管在真核细胞的高尔基体复合物中可以发现有膜性小管延伸并与其相关联,但其生理功能仍不清楚。为了深入了解膜性小管的生物学意义,我们开发了选择性阻止其形成的方法。我们在此表明,多种磷脂酶A2(PLA2)拮抗剂不仅能阻止在高尔基体复合物组装后期发生的膜性小管介导的事件,还能通过诱导其可逆性碎片化成为单独的“小堆叠”来扰乱其正常的稳态微管泡结构。此外,我们表明这些相同的化合物在体外重构系统中可阻止高尔基体堆叠形成膜性小管。该体外测定进一步用于证明相关的PLA2活性源自细胞质。综上所述,这些结果表明,对强效且选择性的PLA2拮抗剂敏感的高尔基体膜性小管介导了高尔基体复合物重新组装过程中的后期事件及其稳态结构的动态维持。此外,这些结果还表明细胞质PLA2酶在介导这些膜运输事件中发挥作用。