San Pietro Enrica, Capestrano Mariagrazia, Polishchuk Elena V, DiPentima Alessio, Trucco Alvar, Zizza Pasquale, Mariggiò Stefania, Pulvirenti Teodoro, Sallese Michele, Tete Stefano, Mironov Alexander A, Leslie Christina C, Corda Daniela, Luini Alberto, Polishchuk Roman S
Department of Cell Biology and Oncology, Consorzio Mario Negri Sud, Chieti, Italy.
PLoS Biol. 2009 Sep;7(9):e1000194. doi: 10.1371/journal.pbio.1000194. Epub 2009 Sep 15.
The organization of intra-Golgi trafficking and the nature of the transport intermediates involved (e.g., vesicles, tubules, or tubular continuities) remain incompletely understood. It was recently shown that successive cisternae in the Golgi stack are interconnected by membrane tubules that form during the arrival of transport carriers from the endoplasmic reticulum. Here, we examine the mechanisms of generation and the function of these tubules. In principle, tubule formation might depend on several protein- and/or lipid-based mechanisms. Among the latter, we have studied the phospholipase A(2) (PLA(2))-mediated generation of wedge-shaped lysolipids, with the resulting local positive membrane curvature. We show that the arrival of cargo at the Golgi complex induces the recruitment of Group IVA Ca(2+)-dependent, cytosolic PLA(2) (cPLA(2)alpha) onto the Golgi complex itself, and that this cPLA(2)alpha is required for the formation of the traffic-dependent intercisternal tubules and for intra-Golgi transport. In contrast, silencing of cPLA(2)alpha has no inhibitory effects on peri-Golgi vesicles. These findings identify cPLA(2)alpha as the first component of the machinery that is responsible for the formation of intercisternal tubular continuities and support a role for these continuities in transport through the Golgi complex.
高尔基体内部运输的组织方式以及所涉及的运输中间体的性质(例如,囊泡、小管或管状连续结构)仍未完全明确。最近有研究表明,高尔基体堆叠中的连续扁平囊泡通过在内质网运输载体到达时形成的膜性小管相互连接。在此,我们研究了这些小管的形成机制及其功能。原则上,小管的形成可能依赖于多种基于蛋白质和/或脂质的机制。在后者中,我们研究了磷脂酶A2(PLA2)介导的楔形溶血磷脂的生成,从而产生局部的正膜曲率。我们发现,货物到达高尔基体复合物会诱导IV型钙离子依赖性胞质磷脂酶A2(cPLA2α)募集到高尔基体复合物本身,并且这种cPLA2α是形成依赖于运输的扁平囊泡间小管以及高尔基体内部运输所必需的。相比之下,沉默cPLA2α对高尔基体周围囊泡没有抑制作用。这些发现确定cPLA2α是负责形成扁平囊泡间管状连续结构的机制中的首个成分,并支持这些连续结构在通过高尔基体复合物的运输中发挥作用。