Differential regulation of sentrinized proteins by a novel sentrin-specific protease
Differential regulation of sentrinized proteins by a novel sentrin-specific protease
复制标题
DOI：
10.1074/jbc.275.5.3355
复制
发表时间：
2000-02-04
影响因子：
4.8
通讯作者：
Yeh, ETH
中科院分区：
文献类型：
--
作者：
Gong, LM;Millas, S;Yeh, ETH
Sentrin-1，又称SUMO-1，是一个由101个氨基酸残基组成的蛋白质，与泛素和另一种泛素样蛋白NEDD 8有较远的亲缘关系。然而，SENP 1与酵母Smt 3特异性蛋白酶Ulp 1关系较远。使用COS细胞表达系统来证明SENP 1的体内活性。当HA标记的sentrin-1与SENP 1共表达时，较高分子量的sentrin-1缀合物被完全去除。令人惊讶的是，在90 kDa的主要sentrinized带保持完整。高分子量sentrin-1缀合物的消失也与游离sentrin-1单体的增加同时发生。SENP 1也对由sentrin-2修饰的蛋白质有活性，但对由泛素或NEDD 8修饰的蛋白质没有活性。此外，sentrinized PML，一种位于细胞核中的肿瘤抑制蛋白，选择性地受到SENP 1的影响，而sentrinized RanGAP 1，与核孔复合物的细胞质纤维相关，保持完整。SENP 1不能在体内处理sentrinized RanGAP 1很可能是由于其核定位，因为SENP 1在体外对sentrinized RanGAP 1有活性。核定位，sentrin特异性蛋白酶的鉴定将提供一个独特的工具，研究sentrin在PML的生物学功能和急性早幼粒细胞白血病的发病机制中的作用。
Sentrin-1, also called SUMO-1, is a protein of 101 residues that is distantly related to ubiquitin and another ubiquitin-like protein, NEDD8, Here we report the cloning of a novel sentrin-specific protease, SENP1, which has no homology to the known de-ubiquitinating enzymes or ubiquitin C-terminal hydrolases. However, SENP1 is distantly related to the yeast Smt3-specific protease, Ulp1. A COS cell expression system was used to demonstrate the activity of SENP1 in vivo. When HA-tagged sentrin-1 was co-expressed with SENP1, the higher molecular weight sentrin-1 conjugates were completely removed. Surprisingly, the major sentrinized band at 90 kDa remained intact. The disappearance of the high molecular weight sentrin-1 conjugates also coincided with an increase in free sentrin-1 monomers. SENP1 is also active against proteins modified by sentrin-2, but not those modified by ubiquitin or NEDD8. In addition, sentrinized PML, a tumor suppressor protein that resides in the nucleus, was selectively affected by SENP1, whereas sentrinized RanGAP1, which is associated with the cytoplasmic fibrils of the nuclear pore complex, remained intact. The inability of SENP1 to process sentrinized RanGAP1 in vivo is most likely due to its nuclear localization because SENP1 is active against sentrinized RanGAP1 in vitro. The identification of a nuclear-localized, sentrin-specific protease will provide a unique tool to study the role of sentrinization in the biological function of PML and in the pathogenesis of acute promyelocytic leukemia.