Arabidopsis TCP Transcription Factors Interact with the SUMO Conjugating Machinery in Nuclear Foci.

Arabidopsis TCP Transcription Factors Interact with the SUMO Conjugating Machinery in Nuclear Foci.
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DOI:
10.3389/fpls.2017.02043
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发表时间:
2017
影响因子:
5.6
通讯作者:
van den Burg HA
van den Burg HA
中科院分区:
生物学2区
文献类型:
--
作者:
Mazur MJ;Spears BJ;Djajasaputra A;van der Gragt M;Vlachakis G;Beerens B;Gassmann W;van den Burg HA

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在拟南芥中,超过400种蛋白质已被确定为SUMO靶点,无论是体内还是体外。其中,转录因子(tf)是SUMO偶联的常见靶标。在这里,我们的目标是使用包含超过1,100个tf的阵列酵母双杂交文库来彻底筛选与SUMO机制相互作用的tf。我们确定了76个相互作用物,它们首先与SUMO偶联酶SCE1和/或SUMO E3连接酶SIZ1相互作用。这些相互作用物属于多种TF家族,它们控制着植物发育和胁迫信号的广泛过程。在这些相互作用物中,TCP家族被过度代表,有几个TCP与SUMO偶联周期的不同蛋白质相互作用。对于这些tcp的一个子集,我们证实了SCE1的催化位点对这种相互作用是必不可少的。与此一致的是,TCP1、TCP3、TCP8、TCP14和TCP15在大肠杆菌SUMO化实验中很容易被SUMO修饰。引人注目的是,这些TCP- sce1相互作用被发现将这些TCP重新分配到核焦点/斑点中,这表明这些TCP焦点代表了SUMO(偶联)活性的位点。
In Arabidopsis more than 400 proteins have been identified as SUMO targets, both in vivo and in vitro. Among others, transcription factors (TFs) are common targets for SUMO conjugation. Here we aimed to exhaustively screen for TFs that interact with the SUMO machinery using an arrayed yeast two-hybrid library containing more than 1,100 TFs. We identified 76 interactors that foremost interact with the SUMO conjugation enzyme SCE1 and/or the SUMO E3 ligase SIZ1. These interactors belong to various TF families, which control a wide range of processes in plant development and stress signaling. Amongst these interactors, the TCP family was overrepresented with several TCPs interacting with different proteins of the SUMO conjugation cycle. For a subset of these TCPs we confirmed that the catalytic site of SCE1 is essential for this interaction. In agreement, TCP1, TCP3, TCP8, TCP14, and TCP15 were readily SUMO modified in an E. coli sumoylation assay. Strikingly, these TCP-SCE1 interactions were found to redistribute these TCPs into nuclear foci/speckles, suggesting that these TCP foci represent sites for SUMO (conjugation) activity.
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