Harrison Rene E, Bucci Cecilia, Vieira Otilia V, Schroer Trina A, Grinstein Sergio
Division of Cell Biology, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada M5G 1X8.
Mol Cell Biol. 2003 Sep;23(18):6494-506. doi: 10.1128/MCB.23.18.6494-6506.2003.
Nascent phagosomes must undergo a series of fusion and fission reactions to acquire the microbicidal properties required for the innate immune response. Here we demonstrate that this maturation process involves the GTPase Rab7. Rab7 recruitment to phagosomes was found to precede and to be essential for their fusion with late endosomes and/or lysosomes. Active Rab7 on the phagosomal membrane associates with the effector protein RILP (Rab7-interacting lysosomal protein), which in turn bridges phagosomes with dynein-dynactin, a microtubule-associated motor complex. The motors not only displace phagosomes in the centripetal direction but, strikingly, promote the extension of phagosomal tubules toward late endocytic compartments. Fusion of tubules with these organelles was documented by fluorescence and electron microscopy. Tubule extension and fusion with late endosomes and/or lysosomes were prevented by expression of a truncated form of RILP lacking the dynein-dynactin-recruiting domain. We conclude that full maturation of phagosomes requires the retrograde emission of tubular extensions, which are generated by activation of Rab7, recruitment of RILP, and consequent association of phagosomes with microtubule-associated motors.
新生吞噬体必须经历一系列融合和裂变反应,以获得先天免疫反应所需的杀菌特性。在此,我们证明这种成熟过程涉及GTP酶Rab7。研究发现,Rab7募集到吞噬体之前,对于吞噬体与晚期内体和/或溶酶体的融合至关重要。吞噬体膜上的活性Rab7与效应蛋白RILP(Rab7相互作用溶酶体蛋白)相关联,而RILP反过来将吞噬体与动力蛋白-动力蛋白激活蛋白连接起来,后者是一种与微管相关的马达复合体。这些马达不仅使吞噬体向心位移,而且惊人的是,促进吞噬体小管向晚期内吞区室延伸。通过荧光和电子显微镜记录了小管与这些细胞器的融合。缺乏动力蛋白-动力蛋白激活蛋白募集结构域的截短形式RILP的表达可阻止小管延伸以及与晚期内体和/或溶酶体的融合。我们得出结论,吞噬体的完全成熟需要通过Rab7激活、RILP募集以及随后吞噬体与微管相关马达的结合来产生管状延伸的逆行排放。