Distinct phylogenetic relationships and biochemical properties of Arabidopsis ovarian tumor-related deubiquitinases support their functional differentiation.
Distinct phylogenetic relationships and biochemical properties of Arabidopsis ovarian tumor-related deubiquitinases support their functional differentiation.
复制标题
DOI：
10.3389/fpls.2014.00084
复制
发表时间：
2014
影响因子：
5.6
通讯作者：
Fu H
中科院分区：
文献类型：
--
作者：
Radjacommare R;Usharani R;Kuo CH;Fu H
泛素化的逆反应由不同种类的去泛素化酶（DUB）催化，包括含有卵巢肿瘤结构域（OTU）的DUB;使用智人蛋白的实验已经证明OTU DUB调节各种细胞过程。除OTLD 1外，植物OTU DUB尚未表征。我们鉴定了12个拟南芥OTU基因座，并在体外分析了11个编码蛋白，以确定它们对M1，K48和K63连接的泛素（UB）链以及UB-/RUB-/SUMO-GST融合的偏好。助理拟南芥OTU DUB显示为半胱氨酸蛋白酶，并分为具有不同连接偏好的四组：OTU 1（M1 = K48 > K63）、OTU 3/4/7/10（K63 > K48 > M1）、OTU 2/9（K48 = K63）和OTU 5/11/12/OTLD 1（无活性）。五个活性OTU DUB（OTU 3/4/7/9/10）也切割RUB融合。OTU 1/3/4切割M1 UB链，表明M1链在植物细胞信号传导中的可能作用。不同A.拟南芥OTU DUBs表明涉及不同的结构元件;例如，OTU 1含氧阴离子残基D89对于裂解异肽键连接的链是必需的，但对于M1链是必需的。仅检测到OTU 2和OTLD 1的UB结合活性，具有不同的连锁偏好。这些生化特性的差异支持了A. thaliana OTU DUBs在不同的功能。此外，根据已建立的系统发育树，植物-和H。存在智人特异性进化枝，这表明这些进化枝中的蛋白质具有分类群特异性功能。我们还检测到5个在物种间保守的OTU进化枝，这表明每个进化枝内不同物种中的直系同源物参与保守的细胞过程，如ERAD和DNA损伤反应。然而，在潜在的跨物种OTU直系同源物中检测到不同的连锁偏好，表明功能和机制分化。
The reverse reaction of ubiquitylation is catalyzed by different classes of deubiquitylation enzymes (DUBs), including ovarian tumor domain (OTU)-containing DUBs; experiments using Homo sapiens proteins have demonstrated that OTU DUBs modulate various cellular processes. With the exception of OTLD1, plant OTU DUBs have not been characterized. We identified 12 Arabidopsis thaliana OTU loci and analyzed 11 of the encoded proteins in vitro to determine their preferences for the ubiquitin (UB) chains of M1, K48, and K63 linkages as well as the UB-/RUB-/SUMO-GST fusions. The A. thaliana OTU DUBs were shown to be cysteine proteases and classified into four groups with distinct linkage preferences: OTU1 (M1 = K48 > K63), OTU3/4/7/10 (K63 > K48 > M1), OTU2/9 (K48 = K63), and OTU5/11/12/OTLD1 (inactive). Five active OTU DUBs (OTU3/4/7/9/10) also cleaved RUB fusion. OTU1/3/4 cleaved M1 UB chains, suggesting a possible role for M1 chains in plant cellular signaling. The different substrate specificities of the various A. thaliana OTU DUBs indicate the involvement of distinct structural elements; for example, the OTU1 oxyanion residue D89 is essential for cleaving isopeptide bond-linked chains but dispensable for M1 chains. UB-binding activities were detected only for OTU2 and OTLD1, with distinct linkage preferences. These differences in biochemical properties support the involvement of A. thaliana OTU DUBs in different functions. Moreover, based on the established phylogenetic tree, plant- and H. sapiens-specific clades exist, which suggests that the proteins within these clades have taxa-specific functions. We also detected five OTU clades that are conserved across species, which suggests that the orthologs in different species within each clade are involved in conserved cellular processes, such as ERAD and DNA damage responses. However, different linkage preferences have been detected among potential cross-species OTU orthologs, indicating functional and mechanistic differentiation.
登录
查看更多内容
影响因子：
14.9
作者：
Block-Schmidt AS;Dukowic-Schulze S;Wanieck K;Reidt W;Puchta H
通讯作者：
Puchta H
影响因子：
5.4
作者：
Fatimababy, Antony S.;Lin, Ya-Ling;Fu, Hongyong
通讯作者：
Fu, Hongyong
影响因子：
7.7
作者：
Balakirev, MY;Tcherniuk, SO;Chroboczek, J
通讯作者：
Chroboczek, J
影响因子：
7.2
作者：
Doelling, JH;Yan, N;Vierstra, RD
通讯作者：
Vierstra, RD
影响因子：
16
作者：
Ernst, Robert;Mueller, Britta;Ploegh, Hidde L.;Schlieker, Christian
通讯作者：
Schlieker, Christian