SUMO targeting of a stress-tolerant Ulp1 SUMO protease.
SUMO targeting of a stress-tolerant Ulp1 SUMO protease.
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SUMO靶向耐应力的ULP1 SUMO蛋白酶。
DOI：
10.1371/journal.pone.0191391
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Kerscher O
中科院分区：
文献类型：
--
作者：
Peek J;Harvey C;Gray D;Rosenberg D;Kolla L;Levy-Myers R;Yin R;McMurry JL;Kerscher O
SENP/Ulp家族的SUMO蛋白酶是SUMO化和去SUMO化的主要调节剂，并调节真核细胞中的SUMO稳态。SUMO结合物响应于细胞应激而迅速增加，包括营养饥饿、缺氧、渗透压应激、DNA损伤、热休克和其他蛋白毒性应激源。然而，对SUMO蛋白酶在应激过程中的调节和靶向知之甚少。为此，我们进行了详细的比较SUMO结合活性的芽殖酵母蛋白Ulp 1（ScUlp 1）和其直系同源物在耐热酵母Kluyveromyces marxianus，KmUlp 1。我们发现ScUlp 1和KmUlp 1的催化UD结构域都显示出高度的序列保守性，在体内补充ulp 1 Δ突变体，并在体外加工SUMO前体。接下来，为了比较两种SUMO蛋白酶的SUMO捕获特征，我们产生了ScUlp 1和KmUlp 1的UD结构域的无催化活性的重组片段，分别称为ScUTAG和KmUTAG。ScUTAG和KmUTAG都能够有效地结合各种纯化的SUMO同种型，并以纳摩尔亲和力结合固定化的SUMO 1。然而，KmUTAG表现出极大增强的能力，结合SUMO和SUMO修饰的蛋白质在氧化，温度和其他应激诱导蛋白质错误折叠的存在下。我们还研究了是否SUMO相互作用的基序（SIM）在UD域的KmULP 1是不保守的ScUlp 1可能有助于SUMO结合特性的KmUTAG。总之，我们的数据揭示了SUMO蛋白酶如何靶向和结合其sumoylated底物的重要细节，特别是在应激条件下。我们还表明，KmUTAG的强大的泛SUMO结合功能，可用于检测和研究细胞培养系统中的SUMO修饰蛋白。
SUMO proteases of the SENP/Ulp family are master regulators of both sumoylation and desumoylation and regulate SUMO homeostasis in eukaryotic cells. SUMO conjugates rapidly increase in response to cellular stress, including nutrient starvation, hypoxia, osmotic stress, DNA damage, heat shock, and other proteotoxic stressors. Nevertheless, little is known about the regulation and targeting of SUMO proteases during stress. To this end we have undertaken a detailed comparison of the SUMO-binding activity of the budding yeast protein Ulp1 (ScUlp1) and its ortholog in the thermotolerant yeast Kluyveromyces marxianus, KmUlp1. We find that the catalytic UD domains of both ScUlp1 and KmUlp1 show a high degree of sequence conservation, complement a ulp1Δ mutant in vivo, and process a SUMO precursor in vitro. Next, to compare the SUMO-trapping features of both SUMO proteases we produced catalytically inactive recombinant fragments of the UD domains of ScUlp1 and KmUlp1, termed ScUTAG and KmUTAG respectively. Both ScUTAG and KmUTAG were able to efficiently bind a variety of purified SUMO isoforms and bound immobilized SUMO1 with nanomolar affinity. However, KmUTAG showed a greatly enhanced ability to bind SUMO and SUMO-modified proteins in the presence of oxidative, temperature and other stressors that induce protein misfolding. We also investigated whether a SUMO-interacting motif (SIM) in the UD domain of KmULP1 that is not conserved in ScUlp1 may contribute to the SUMO-binding properties of KmUTAG. In summary, our data reveal important details about how SUMO proteases target and bind their sumoylated substrates, especially under stress conditions. We also show that the robust pan-SUMO binding features of KmUTAG can be exploited to detect and study SUMO-modified proteins in cell culture systems.
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DOI：
10.1083/jcb.200212052
发表时间：
2003-03-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Li SJ;Hochstrasser M
通讯作者：
Hochstrasser M
影响因子：
7.8
作者：
Makhnevych, Taras;Ptak, Christopher;Lusk, C Patrick;Aitchison, John D;Wozniak, Richard W
通讯作者：
Wozniak, Richard W
影响因子：
12.3
作者：
Nayak A;Müller S
通讯作者：
Müller S
影响因子：
5.4
作者：
Elmore ZC;Donaher M;Matson BC;Murphy H;Westerbeck JW;Kerscher O
通讯作者：
Kerscher O
影响因子：
3.7
作者：
Boudreau É;Labib S;Bertrand AT;Decostre V;Bolongo PM;Sylvius N;Bonne G;Tesson F
通讯作者：
Tesson F