A novel gene, MdSSK1, as a component of the SCF complex rather than MdSBP1 can mediate the ubiquitination of S-RNase in apple.
A novel gene, MdSSK1, as a component of the SCF complex rather than MdSBP1 can mediate the ubiquitination of S-RNase in apple.
复制标题
苹果中一种新的基因 MdSSK1（而不是 MdSBP1）作为 SCF 复合体的组成部分可以介导 S-RNase 的泛素化。
DOI：
10.1093/jxb/eru164
复制
发表时间：
2014-07
影响因子：
6.9
通讯作者：
Li T
中科院分区：
文献类型：
--
作者：
Yuan H;Meng D;Gu Z;Li W;Wang A;Yang Q;Zhu Y;Li T
克隆了苹果SCF复合物的相关组分，证明含有MdSSK 1而不是MdSBP 1的SCF复合物可以介导S-RNase的泛素化。SCF（SKP 1-Cullin 1-F-box-Rbx 1）复合体（包括花粉决定子SLF，S-locus-F-box）作为S-RNase为基础的配子体自交不亲和性（GSI）的核心因子，在非自身S-RNase上起E3泛素连接酶的作用。SCF复合物由SKP 1桥接SLF、CUL 1和Rbx 1形成;然而，尚不清楚缺乏SKP 1的SCF复合物是否可以介导S-RNase的泛素化。根据“金冠”苹果基因组结构特征，从花粉中克隆了3个类SKP 1基因。这些基因在C末端有一个标准的“WAFE”后的5个氨基酸基序，此外，还含有8个折叠和2个螺旋。这三个基因都只在花粉中表达。在酵母双杂交和pull-down分析中，只发现一个与MdSFBB和MdCUL 1相互作用的基因，表明该基因是苹果中与SLF相互作用的类SKP 1基因，命名为MdSSK 1。利用MdSSK 1、S2-MdSFBB 1（S2-MalusaparticaS-locus-F-box brother）和MdCUL 1蛋白与S2-RNase和泛素共同孵育的体外实验表明，SCF复合物在体外泛素化S-RNase，而MdSBP 1（MalusaparticaS-RNasebindingprotein 1）不能在功能上取代SCF复合物中的MdSSK 1来泛素化S-RNase。根据上述实验，MdSBP 1可能是负责与S-RNase识别的唯一因子，而不是SCF复合物的组分，并且需要含有MdSSK 1的SCF复合物来介导S-RNase的泛素化。
The related components of the SCF complex in apple were cloned and it was proved that an SCF complex containing MdSSK1 rather than MdSBP1 can mediate the ubiquitination of S-RNase. As a core factor in S-RNase-based gametophytic self-incompatibility (GSI), the SCF (SKP1–Cullin1–F-box-Rbx1) complex (including pollen determinant SLF, S-locus-F-box) functions as an E3 ubiquitin ligase on non-self S-RNase. The SCF complex is formed by SKP1 bridging between SLF, CUL1, and Rbx1; however, it is not known whether an SCF complex lacking SKP1 can mediate the ubiquitination of S-RNase. Three SKP1-like genes from pollen were cloned based on the structural features of the SLF-interacting-SKP1-like (SSK) gene and the ‘Golden Delicious’ apple genome. These genes have a motif of five amino acids following the standard ‘WAFE’ at the C terminal and, in addition, contain eight sheets and two helices. All three genes were expressed exclusively in pollen. In the yeast two-hybrid and pull-down assays only one was found to interact with MdSFBB and MdCUL1, suggesting it is the SLF-interacting SKP1-like gene in apple which was named MdSSK1. In vitro experiments using MdSSK1, S2-MdSFBB1 (S2-Malus domestica S-locus-F-box brother) and MdCUL1 proteins incubated with S 2-RNase and ubiquitin revealed that the SCF complex ubiquitinylates S-RNase in vitro, while MdSBP1 (Malus domestica S-RNase binding protein 1) could not functionally replace MdSSK1 in the SCF complex in ubiquitinylating S-RNase. According to the above experiments, MdSBP1 is probably the only factor responsible for recognition with S-RNase, while not a component of the SCF complex, and an SCF complex containing MdSSK1 is required for mediating the ubiquitination of S-RNase.
登录
查看更多内容
影响因子：
3.4
作者：
Marín I
通讯作者：
Marín I
DOI：
10.1042/bj20081659
发表时间：
2009-05-13
期刊：
The Biochemical journal
影响因子：
--
作者：
Jourdain I;Spielewoy N;Thompson J;Dhut S;Yates JR;Toda T
通讯作者：
Toda T
DOI：
10.1073/pnas.162339999
发表时间：
2002-08-20
影响因子：
11.1
作者：
Gagne, JM;Downes, BP;Vierstra, RD
通讯作者：
Vierstra, RD
影响因子：
11.6
作者：
Hua, Zhihua;Kao, Teh-hui
通讯作者：
Kao, Teh-hui
影响因子：
4.9
作者：
Chang, Liang-Chi;Guo, Cian-Ling;Jauh, Guang-Yuh
通讯作者：
Jauh, Guang-Yuh