A Yeast-Based Functional Assay to Study Plant N-Degron - N-Recognin Interactions.
A Yeast-Based Functional Assay to Study Plant N-Degron - N-Recognin Interactions.
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DOI：
10.3389/fpls.2021.806129
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发表时间：
2021
影响因子：
5.6
通讯作者：
Bachmair A
中科院分区：
文献类型：
--
作者：
Kozlic A;Winter N;Telser T;Reimann J;Rose K;Nehlin L;Berckhan S;Sharma G;Dambire C;Boeckx T;Holdsworth MJ;Bachmair A
N-degron途径是泛素-蛋白酶体系统的一个分支，其中氨基末端残基作为降解信号。通过合成生物学方法，我们将拟南芥中的泛素连接酶PRT6和泛素偶联酶2 （AtUBC2）表达到内源性N-degron通路突变的酿酒酵母菌株中。这两种酶重新构成了植物N-degron途径的一部分，并通过监测从精氨酸N-degrons开始的共表达gfp连接的植物蛋白的稳定性来探测。新的分析方法允许直接分析，而体外相互作用分析通常不允许检测N-degron识别泛素连接酶与其底物的弱结合，并且在植物中测试通常是复杂和耗时的。
The N-degron pathway is a branch of the ubiquitin-proteasome system where amino-terminal residues serve as degradation signals. In a synthetic biology approach, we expressed ubiquitin ligase PRT6 and ubiquitin conjugating enzyme 2 (AtUBC2) from Arabidopsis thaliana in a Saccharomyces cerevisiae strain with mutation in its endogenous N-degron pathway. The two enzymes re-constitute part of the plant N-degron pathway and were probed by monitoring the stability of co-expressed GFP-linked plant proteins starting with Arginine N-degrons. The novel assay allows for straightforward analysis, whereas in vitro interaction assays often do not allow detection of the weak binding of N-degron recognizing ubiquitin ligases to their substrates, and in planta testing is usually complex and time-consuming.
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影响因子：
5.3
作者：
Ramadan A;Nemoto K;Seki M;Shinozaki K;Takeda H;Takahashi H;Sawasaki T
通讯作者：
Sawasaki T
影响因子：
7.4
作者：
Zhang, Hongtao;Linster, Eric;Wirtz, Markus
通讯作者：
Wirtz, Markus
影响因子：
2.6
作者：
WACH, A;BRACHAT, A;PHILIPPSEN, P
通讯作者：
PHILIPPSEN, P
影响因子：
56.9
作者：
BACHMAIR, A;FINLEY, D;VARSHAVSKY, A
通讯作者：
VARSHAVSKY, A
影响因子：
16.6
作者：
White MD;Klecker M;Hopkinson RJ;Weits DA;Mueller C;Naumann C;O'Neill R;Wickens J;Yang J;Brooks-Bartlett JC;Garman EF;Grossmann TN;Dissmeyer N;Flashman E
通讯作者：
Flashman E