The membrane-tethering protein p115 interacts with GBF1, an ARF guanine-nucleotide-exchange factor
The membrane-tethering protein p115 interacts with GBF1, an ARF guanine-nucleotide-exchange factor
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DOI：
10.1038/sj.embor.embor762
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发表时间：
2003-03-01
期刊：
影响因子：
7.7
通讯作者：
Sztul, E
中科院分区：
文献类型：
--
作者：
García-Mata, R;Sztul, E
膜转运因子p115与膜转运机制的不同组分相互作用。它结合两种高尔基体基质蛋白、一种Rab GT3和可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体（SNARE）家族的各种成员。在这里，我们描述了一种新的p115和高尔基体特异性布雷菲德-A-抗性因子1（GBF 1），鸟嘌呤核苷酸交换因子ADP核糖基化因子（ARF）之间的相互作用。在酵母双杂交筛选中，使用全长p115作为诱饵鉴定了GBF 1。使用体外和体内试验，在生物化学上证实了相互作用。相互作用结构域被映射到GBF 1的富含脯氨酸区域和p115的头部区域。这些蛋白质广泛地共定位在高尔基体和外周泡状小管簇中。突变分析表明，相互作用不是将GBF 1或p115靶向细胞膜所必需的。GBF 1的p115结合（富含亲）区域的表达导致高尔基体破坏，表明p115和GBF 1之间的相互作用是功能相关的。
The membrane-transport factor p115 interacts with diverse components of the membrane-transport machinery. It binds two Golgi matrix proteins, a Rab GTPase, and various members of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) family. Here, we describe a novel interaction between p115 and Golgi-specific brefeldin-A-resistant factor 1 (GBF1), a guanine-nucleotide exchange factor for ADP ribosylation factor (ARF). GBF1 was identified in a yeast two-hybrid screen, using full-length p115 as bait. The interaction was confirmed biochemically, using in vitro and in vivo assays. The interacting domains were mapped to the proline-rich region of GBF1 and the head region of p115. These proteins colocalize extensively in the Golgi and in peripheral vesicular tubular clusters. Mutagenesis analysis indicates that the interaction is not required for targeting GBF1 or p115 to membranes. Expression of the p115-binding (pro-rich) region of GBF1 leads to Golgi disruption, indicating that the interaction between p115 and GBF1 is functionally relevant.