Gillingham Alison K, Tong Amy Hin Yan, Boone Charles, Munro Sean
Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, England, UK.
J Cell Biol. 2004 Oct 25;167(2):281-92. doi: 10.1083/jcb.200407088.
Rud3p is a coiled-coil protein of the yeast cis-Golgi. We find that Rud3p is localized to the Golgi via a COOH-terminal domain that is distantly related to the GRIP domain that recruits several coiled-coil proteins to the trans-Golgi by binding the small Arf-like GTPase Arl1p. In contrast, Rud3p binds to the GTPase Arf1p via this COOH-terminal "GRIP-related Arf-binding" (GRAB) domain. Deletion of RUD3 is lethal in the absence of the Golgi GTPase Ypt6p, and a screen of other mutants showing a similar genetic interaction revealed that Golgi targeting of Rud3p also requires Erv14p, a cargo receptor that cycles between the endoplasmic reticulum and Golgi. The one human protein with a GRAB domain, GMAP-210 (CEV14/Trip11/Trip230), is known to be on the cis-Golgi, but the COOH-terminal region that contains the GRAB domain has been reported to bind to centrosomes and gamma-tubulin (Rios, R.M, A. Sanchis, A.M. Tassin, C. Fedriani, and M. Bornens. 2004. Cell. 118:323-335). In contrast, we find that this region binds to the Golgi in a GRAB domain-dependent manner, suggesting that GMAP-210 may not link the Golgi to gamma-tubulin and centrosomes.
Rud3p是酵母顺式高尔基体中的一种卷曲螺旋蛋白。我们发现，Rud3p通过其COOH末端结构域定位于高尔基体，该结构域与GRIP结构域有较远的亲缘关系，GRIP结构域通过结合小的Arf样GTP酶Arl1p将几种卷曲螺旋蛋白招募到反式高尔基体。相反，Rud3p通过这个COOH末端的“GRIP相关Arf结合”（GRAB）结构域与GTP酶Arf1p结合。在没有高尔基体GTP酶Ypt6p的情况下，RUD3的缺失是致死性的，对其他显示类似遗传相互作用的突变体进行筛选发现，Rud3p的高尔基体靶向也需要Erv14p，一种在内质网和高尔基体之间循环的货物受体。已知一种具有GRAB结构域的人类蛋白GMAP-210（CEV14/Trip11/Trip230）位于顺式高尔基体上，但据报道，包含GRAB结构域的COOH末端区域与中心体和γ-微管蛋白结合（Rios, R.M, A. Sanchis, A.M. Tassin, C. Fedriani, and M. Bornens. 2004. Cell. 118:323 - 335）。相比之下，我们发现该区域以GRAB结构域依赖的方式与高尔基体结合，这表明GMAP-210可能不会将高尔基体与γ-微管蛋白和中心体联系起来。
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