Interactions and CCAAT-binding of Arabidopsis thaliana NF-Y subunits.
Interactions and CCAAT-binding of Arabidopsis thaliana NF-Y subunits.
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DOI：
10.1371/journal.pone.0042902
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Tonelli C
中科院分区：
文献类型：
--
作者：
Calvenzani V;Testoni B;Gusmaroli G;Lorenzo M;Gnesutta N;Petroni K;Mantovani R;Tonelli C
核因子-Y是一种具有高度特异性和亲和力的转录因子，能识别广泛存在的CCAAT盒启动子元件。它由三个亚基组成：NF-YA和含组蛋白折叠结构域(HFDs)的NF-YB/NF-YC-杂二聚体。我们以前在拟南芥中发现了一个由29个成员组成的大的NF-Y基因家族，并研究了它们在不同植物组织中的表达模式。我们利用酵母双杂交试验(Y2H)、下拉试验和凝胶迁移试验(EMSA)对重组蛋白进行了体外实验，分析了AtNF-YB/AtNF-YC与不同AtNF-YA的相互作用和与DNA的结合。与HFDs内的保守一致，我们表明所有组蛋白样亚单位之间都可能发生异源二聚化，包括胚胎发育所需的分化和相关的LEC1/AtNF-YB9和L1L/AtNF-YB6。用特定的AtNF-YB/AtNF-YC组合研究了DNA与共识CCAAT盒的结合，并观察到了一些但不是所有AtNF-YA亚基的结合。我们的结果强调了(I)AtNF-Y组蛋白样亚单位保守的异源二聚能力，以及(Ii)AtNF-YAS与CCAAT序列的不同亲和力。由于核因子-Y基因在植物中的普遍扩增，这些结果很可能适用于其他物种。
NF-Y is a transcription factor that recognizes with high specificity and affinity the widespread CCAAT box promoter element. It is formed by three subunits: NF-YA and the NF-YB/NF-YC- heterodimer containing histone fold domains (HFDs). We previously identified a large NF-Y gene family in Arabidopsis thaliana, composed of 29 members, and characterized their expression patterns in various plant tissues. We used yeast Two-hybrids assays (Y2H), pull-down and Electrophoretic Mobility Shift Assay (EMSA) in vitro experiments with recombinant proteins to dissect AtNF-YB/AtNF-YC interactions and DNA-binding with different AtNF-YAs. Consistent with robust conservation within HFDs, we show that heterodimerization is possible among all histone-like subunits, including the divergent and related LEC1/AtNF-YB9 and L1L/AtNF-YB6 required for embryo development. DNA-binding to a consensus CCAAT box was investigated with specific AtNF-YB/AtNF-YC combinations and observed with some, but not all AtNF-YA subunits. Our results highlight (i) the conserved heterodimerization capacity of AtNF-Y histone-like subunits, and (ii) the different affinities of AtNF-YAs for the CCAAT sequence. Because of the general expansion of NF-Y genes in plants, these results most likely apply to other species.
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DOI：
10.1073/pnas.0437909100
发表时间：
2003-02-18
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
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影响因子：
14.9
作者：
LI, XY;MANTOVANI, R;MATHIS, D
通讯作者：
MATHIS, D