The Ulp1 SUMO isopeptidase: distinct domains required for viability, nuclear envelope localization, and substrate specificity.
The Ulp1 SUMO isopeptidase: distinct domains required for viability, nuclear envelope localization, and substrate specificity.
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DOI：
10.1083/jcb.200212052
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发表时间：
2003-03-31
期刊：
影响因子：
--
通讯作者：
Hochstrasser M
中科院分区：
文献类型：
--
作者：
Li SJ;Hochstrasser M
泛素样SUMO蛋白的蛋白质修饰有助于许多细胞调节机制。在酿酒酵母中，sumoylating和desumoylating活动是必不可少的生存力。在其两种已知的去小糖基化酶中，Ubl特异性蛋白酶（Ulp）1和Ulp 2/Smt 4，Ulp 1是细胞周期进程特异性所需的。一个约200个残基的片段，Ulp结构域（UD），在Ulp中是保守的，并且包括一个甚至更广泛的核心半胱氨酸蛋白酶结构域。在这里，我们证明了Ulp 1 UD本身可以支持野生型生长速率，并在体外可以从底物上切割SUMO。然而，在仅表达Ulp 1的UD的细胞中，许多SUMO缀合物积累到高水平，表明非必需的Ulp 1 NH 2-末端结构域对于针对大部分SUMO化靶标的活性是重要的。NH 2-末端结构域还包括在核膜位点浓缩Ulp 1所必需和足够的序列。值得注意的是，与全长Ulp 1不同，NH 2末端缺失的Ulp 1变体能够抑制缺乏分歧Ulp 2异肽酶的细胞的缺陷。因此，Ulp 1的NH 2-末端调节结构域限制了Ulp 1对某些sumoylated蛋白的活性，同时使其他蛋白能够裂解。这些数据定义了Ulp 1的关键功能元件，并强烈表明亚细胞定位是SUMO异肽酶特异性的生理学重要约束。
Protein modification by the ubiquitin-like SUMO protein contributes to many cellular regulatory mechanisms. In Saccharomyces cerevisiae, both sumoylating and desumoylating activities are essential for viability. Of its two known desumoylating enzymes, Ubl-specific protease (Ulp)1 and Ulp2/Smt4, Ulp1 is specifically required for cell cycle progression. A ∼200-residue segment, the Ulp domain (UD), is conserved among Ulps and includes a core cysteine protease domain that is even more widespread. Here we demonstrate that the Ulp1 UD by itself can support wild-type growth rates and in vitro can cleave SUMO from substrates. However, in cells expressing only the UD of Ulp1, many SUMO conjugates accumulate to high levels, indicating that the nonessential Ulp1 NH2-terminal domain is important for activity against a substantial fraction of sumoylated targets. The NH2-terminal domain also includes sequences necessary and sufficient to concentrate Ulp1 at nuclear envelope sites. Remarkably, NH2-terminally deleted Ulp1 variants are able, unlike full-length Ulp1, to suppress defects of cells lacking the divergent Ulp2 isopeptidase. Thus, the NH2-terminal regulatory domain of Ulp1 restricts Ulp1 activity toward certain sumoylated proteins while enabling the cleavage of others. These data define key functional elements of Ulp1 and strongly suggest that subcellular localization is a physiologically significant constraint on SUMO isopeptidase specificity.
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