Identification and characterization of NEDD8-modified proteins from plants

NEDD 8是真核细胞中的泛素相关蛋白修饰剂。Neddylation调节动物、植物和真菌中cullin-RING型E3泛素连接酶的组装和活性。除了cullin，已知的NEDD 8修饰蛋白的数量非常有限，但还没有从植物中报道过这样的蛋白。使用标记的版本的拟南芥NEDD 8（RUB 1），我们已经纯化了一些推定的NEDD 8共轭植物模式物种拟南芥。在这里，我们建议在体内确认这些蛋白质的neddylation，映射的neddylation网站，并检查neddylation蛋白质功能的相关性。我们还建议建立一个更精细的纯化方案NEDD 8缀合物得到一个更完整的概述植物NEDD 8 ylome和检查NEDD 8修饰的蛋白质的差异积累在NEDD 8缀合和去缀合缺陷的突变体。

NEDD8 is a ubiquitin-related protein modifier in eukaryotic cells. Neddylation regulates the assembly and activity of cullin-RING type E3 ubiquitin ligases in animals, plants, and fungi. Besides cullins, only a very limited number of NEDD8-modified proteins are known, but no such proteins have as yet been reported from plants. Using a tagged version of Arabidopsis NEDD8 (RUB1), we have purified a number of putative NEDD8-conjugates from the plant model species Arabidopsis thaliana. Here, we propose to confirm the neddylation of these proteins in vivo, to map the neddylation sites, and to examine the relevance of neddylation for protein function. We also propose to establish a more refined purification scheme for NEDD8 conjugates to get a more complete overview of the plant NEDD8ylome and to examine the differential accumulation of NEDD8-modified proteins in mutants deficient in NEDD8 conjugation and deconjugation.