DEN1 deneddylates non-cullin proteins in vivo
DEN1 deneddylates non-cullin proteins in vivo
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DOI：
10.1242/jcs.030445
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发表时间：
2008-10-01
影响因子：
4
通讯作者：
Chien, Cheng-Ting
中科院分区：
文献类型：
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作者：
Chan, Yaru;Yoon, Jeongsook;Chien, Cheng-Ting
泛素样蛋白Nedd 8/Rub 1共价修饰并激活cullin泛素连接酶。然而，Nedd 8修饰的蛋白质的库和蛋白质neddylation状态的调节尚不清楚。半胱氨酸蛋白酶DEN 1/NEDP 1特异性地加工Nedd 8前体，并且已经被建议将Nedd 8与cullin蛋白解缀合。通过表征果蝇DEN 1蛋白和DEN 1空（DEN 1（空））突变体，我们提供了在体外和体内的证据，DEN 1，除了处理Nedd 8，deneddylates许多细胞蛋白。尽管纯化的DEN 1蛋白有效地去涡化Nedd 8-缀合的cullin蛋白Cul 1和Cul 3，但去涡化Cul 1和Cul 3蛋白水平在DEN 1（无效）中没有增强。引人注目的是，许多细胞蛋白在DEN 1突变体中高度neddylated，并被纯化的DEN 1蛋白去eddylated。DEN 1去螺旋化活性与cullin去螺旋化CSN的活性不同。遗传分析表明，由DEN 1维持的neddylation和deneddylation之间的平衡对动物的生存能力至关重要。
The ubiquitin-like protein Nedd8/Rub1 covalently modifies and activates cullin ubiquitin ligases. However, the repertoire of Nedd8-modified proteins and the regulation of protein neddylation status are not clear. The cysteine protease DEN1/NEDP1 specifically processes the Nedd8 precursor and has been suggested to deconjugate Nedd8 from cullin proteins. By characterizing the Drosophila DEN1 protein and DEN1 null (DEN1(null)) mutants, we provide in vitro and in vivo evidence that DEN1, in addition to processing Nedd8, deneddylates many cellular proteins. Although purified DEN1 protein efficiently deneddylates the Nedd8-conjugated cullin proteins Cul1 and Cul3, neddylated Cul1 and Cul3 protein levels are not enhanced in DEN1(null). Strikingly, many cellular proteins are highly neddylated in DEN1 mutants and are deneddylated by purified DEN1 protein. DEN1 deneddylation activity is distinct from that of the cullin-deneddylating CSN. Genetic analyses indicate that a balance between neddylation and deneddylation maintained by DEN1 is crucial for animal viability.