Identification of ASK and clock-associated proteins as molecular partners of LKP2 (LOV kelch protein 2) in Arabidopsis.
Identification of ASK and clock-associated proteins as molecular partners of LKP2 (LOV kelch protein 2) in Arabidopsis.
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DOI：
10.1093/jxb/erh226
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发表时间：
2004-09
影响因子：
6.9
通讯作者：
M. Yasuhara;Shunya Mitsui;H. Hirano;Rieko Takanabe;Y. Tokioka;N. Ihara;Akihiro Komatsu;M. Seki;K. Shinozaki;T. Kiyosue
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文献类型：
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作者：
M. Yasuhara;Shunya Mitsui;H. Hirano;Rieko Takanabe;Y. Tokioka;N. Ihara;Akihiro Komatsu;M. Seki;K. Shinozaki;T. Kiyosue
拟南芥（Arabidopsis thaliana Heynh.）具有LOV结构域、F-box基序和kelch重复区。LKP 2是该家族的成员，在拟南芥中在昼夜节律振荡器内或非常接近昼夜节律振荡器发挥作用。启动子-GUS融合研究表明，LKP 2基因在莲座叶中高度活跃。在CaMV 35 S：LKP 2-GFP植物中，在细胞核中检测到GFP相关的荧光，表明LKP 2是一种核蛋白。酵母双杂交分析表明，LKP 2与拟南芥Skp 1-like蛋白（ASK）以及其它ADO/FKF/LKP/ZTL家族蛋白相互作用，表明LKP 2可以形成SCF（Skp 1-Cullin-F-box protein）复合物，其功能类似于泛素E3连接酶。LKP 2不仅与自身相互作用，还与家族的其他成员LKP 1和FKF 1相互作用。双杂交分析还表明，LKP 2与TOC 1，一个时钟组件，但不与CCA 1或LHY，TOC 1基因表达的负调控。LKP 2的LOV结构域被证明是与TOC 1相互作用所必需和充分的。还观察到LKP 2与TOC 1的旁系同源物APRR 5之间的相互作用，但LKP 2不与TOC 1的其他旁系同源物APRR 3、APRR 7或APRR 9相互作用。
The ADO/FKF/LKP/ZTL family of proteins of Arabidopsis thaliana Heynh. have a LOV domain, an F-box motif, and a kelch repeat region. LKP2 is a member of this family and functions either within or very close to the circadian oscillator in Arabidopsis. Promoter-GUS fusion studies revealed that the LKP2 gene was highly active in rosette leaves. In CaMV 35S:LKP2-GFP plants, GFP-associated fluorescence was detected in nuclei, suggesting that LKP2 is a nuclear protein. Yeast two-hybrid analysis demonstrated that LKP2 interacted with some Arabidopsis Skp1-like proteins (ASK), as do other ADO/FKF/LKP/ZTL family proteins, suggesting that LKP2 can form an SCF (Skp1-Cullin-F-box protein) complex that functions as a ubiquitin E3 ligase. LKP2 interacted not only with itself but also with other members of the family, LKP1 and FKF1. The two-hybrid analysis also demonstrated that LKP2 interacted with TOC1, a clock component, but not with CCA1 or LHY, negative regulators of TOC1 gene expression. The LOV domain of LKP2 was shown to be necessary and sufficient for the interaction with TOC1. An interaction between LKP2 and APRR5, a paralogue of TOC1, was also observed, but LKP2 did not interact with APRR3, APRR7, or APRR9, other paralogues of TOC1.