兰州大学机构库 >生命科学学院
TCP1直接结合启动子区域GGNCCC序列调控DWF4及其他拟南芥基因的表达
Alternative TitleTCP1 modulates the expressions of DWF4 and other genes via directly interacting with the GGNCCC motifs in their promoter regions in Arabidopsis thaliana
高亚虎
Thesis Advisor黎家
2015-06-03
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword油菜素内酯 TCP1 DWF4 拟南芥 体内平衡
AbstractTCP1属于一类植物特有的转录因子家族,称为TCP家族,其家族成员蛋白都包括一段保守的包括59个氨基酸残基的特殊bHLH结构域,叫做TCP功能域,这段序列与DNA结合或者与蛋白质相互作用有关。TCP1属于亚家族II,GTGGNCCC序列是这个家族成员偏好的结合序列,但是在DWF4的启动子中并没有发现与此完全相同的序列,只发现两个GGNCCC核心序列。染色质免疫共沉淀实验结果显示这两个序列可以与TCP1存在相互作用,凝胶迁移速率分析进一步证明这两个序列可以与TCP1直接结合,利用点突变技术证明了此核心序列两侧的其它序列对TCP1的结合并不重要。同时,利用启动子-GUS转基因技术,我们对DWF4启动子中两个GGNCCC核心序列的重要性进行了研究,与相同长度的野生型启动子相比,当两个核心序列被替换后,启动子的转录效率大大降低了。因为tcp1-1D及TCP1-SRDX植株还表现出一些与BR生物合成及信号途径无关的表型,说明TCP1应该还调控别的与植物生长发育有关的基因,我们于是利用基因芯片技术比较了野生型、TCP1-SRDX及tcp1-1D中全基因组表达图谱的差异,找到了520个能被TCP1-SRDX或tcp1-1D显著调控且在它们的启动子中含有至少一个GGNCCC序列的基因(与野生型比较,这520个基因至少有2倍以上的表达差异),我们挑选了部分表达差异明显的基因进行了PCR验证,证明了芯片结果的可靠性。除了调控DWF4的表达外,TCP1还通过调控其他启动子中含有GGNCCC的基因的表达从而实现对植物生长发育的总体调控。这些实验数据说明TCP1是通过直接结合到GGNCCC序列来发挥调控基因表达功能的。
Other AbstractTCP1 belongs to TCP family, a unique plant transcription factor family, TCP genes encoding proteins containing the so-called TCP domain, a conserved 59-amino acid basic helix–loop–helix (bHLH) motif that allows DNA binding and protein–protein interactions. TCP1 is a member of class II. GTGGNCCC motif was considered as a putative binding motif for TCP1. There is no such a sequence in the promoter region of DWF4. Instead, there are two GGNCCC motifs. ChIP analyses indiated that these two GGNCCC motifs can asssociate with TCP1. Analyses using EMSA further comfirmed that TCP1 can directly interact with the two GGNCCC motifs in the promoter region of DWF4. Site-directed mutagenesis suggested the flanking sequences of GGNCCC are less important for TCP1 binding. At the same time, we analyzed the importance of these two motifs in TCP1 controlled DWF4 expression by using a promoter-GUS transformation technique. Comparing to the same length of native DWF4 promoter, the modified promoter with two TCP1 motifs being replaced showed dramatically decreased transcription efficience. Because overexpression of TCP1 or TCP1-SRDX also showed phenotypes unrelated to BR biosynthesis or signaling, we carried out microarray analyses to compare the transcriptions of genes from wild-type, TCP1-SRDX and tcp1-1D plants, respectively in order to idenfy all potential TCP1-regulated genes in the Arabidopsis genome. We identified 520 genes in which there is at least one GGNCCC motif per gene and their expression levels are up- or down-regulated by at least two folds. These studies suggested that TCP1 not only regulate the expression of the key BR biosynthesis gene DWF4, but also other genes critical to plant growth and development.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221847
Collection生命科学学院
Recommended Citation
GB/T 7714
高亚虎. TCP1直接结合启动子区域GGNCCC序列调控DWF4及其他拟南芥基因的表达[D]. 兰州. 兰州大学,2015.
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