University of Osnabrück, Department of Biology and Chemistry, Biochemistry section, Barbarastrasse 13, 49076 Osnabrück, Germany.
J Cell Sci. 2010 Dec 1;123(Pt 23):4085-94. doi: 10.1242/jcs.071977. Epub 2010 Nov 9.
Organelles of the endomembrane system need to counterbalance fission and fusion events to maintain their surface-to-volume ratio. At the late mammalian endosome, the Rab GTPase Rab7 is a major regulator of fusion, whereas the homologous yeast protein Ypt7 seems to be restricted to the vacuole surface. Here, we present evidence that Ypt7 is recruited to and acts on late endosomes, where it affects multiple trafficking reactions. We show that overexpression of Ypt7 results in expansion and massive invagination of the vacuolar membrane, which requires cycling of Ypt7 between GDP- and GTP-bound states. Invaginations are blocked by ESCRT, CORVET and retromer mutants, but not by autophagy or AP-3 mutants. We also show that Ypt7-GTP specifically binds to the retromer cargo-recognition subcomplex, which--like its cargo Vps10--is found on the vacuole upon Ypt7 overproduction. Our data suggest that Ypt7 functions at the late endosome to coordinate retromer-mediated recycling with the fusion of late endosomes with vacuoles.
内质网膜系统的细胞器需要平衡分裂和融合事件,以维持其表面积与体积的比例。在哺乳动物晚期内体中,Rab GTP 酶 Rab7 是融合的主要调节剂,而同源的酵母蛋白 Ypt7 似乎仅限于液泡表面。在这里,我们提供的证据表明,Ypt7 被招募到晚期内体并在其上发挥作用,影响多种运输反应。我们表明,Ypt7 的过表达导致液泡膜的扩张和大量内陷,这需要 Ypt7 在 GDP 和 GTP 结合状态之间循环。内陷被 ESCRT、CORVET 和逆行转运体突变体阻断,但不是自噬或 AP-3 突变体。我们还表明,Ypt7-GTP 特异性结合逆行转运体货物识别亚基,该亚基(与其货物 Vps10 一样)在 Ypt7 过表达时存在于液泡上。我们的数据表明,Ypt7 在晚期内体中发挥作用,协调逆行转运体介导的循环与晚期内体与液泡的融合。