白花柽柳微小染色体维系蛋白3(TaMCM3)特异性识别W-box元件  

Analysis on Minichromosome Maintenance 3 Protein from Tamarix androssowii Binding to W-box Element

作　　者：刘文进[1] 王留强[1] 王玉成[1] 

机构地区：[1]林木遗传育种国家重点实验室(东北林业大学),哈尔滨150040

出　　处：《东北林业大学学报》2013年第8期22-26,共5页Journal of Northeast Forestry University

基　　金：全国优秀博士学位论文作者专项基金资助(200973)

摘　　要：真核生物翻译起始因子5A(eukaryotic translation initiation factor 5A,eIF5A)是一个普遍存在于真核生物中高度保守的蛋白,参与多种生命活动。通过对已克隆得到的一个柽柳eIF5A基因(TaeIF5A1)启动子分析发现,其-622～-627 bp区域存在W-box"CTGACT"元件,利用酵母单杂交技术筛选柽柳cDNA文库获得一个能够识别W-box的微小染色体维系蛋白3(minichromosome maintenance 3,MCM3),命名为TaMCM3。进一步利用酵母单杂交验证发现,将W-box的核心序列"TGAC"中的"A"突变成"G"或者将"G"突变成"A",TaMCM则不能识别这些序列,说明TaMCM3对W-box的识别具有特异性。TaMCM能够结合含有W-box元件的TaeIF5A1启动子片段,但当W-box序列突变后则不能识别,说明TaMCM是通过与W-box相结合来与TaeIF5A1启动子相互作用的。研究表明,TaMCM能够特异性地识别W-box,并可能具有转录因子的功能。Eukaryotic translation initiation factor 5A （ eIFSA）, a group of highly conserved proteins, are distributed in eukaryot- ic cells and involved in a variety of biological processes. A W-box ＂CTGACT＂ motif was in -622-627 bp in the promoter of an elFSA gene （ TaelFSA1 ） from Tamarix androssowii. A mini-chromosome maintenance protein （ denoted as TaMCM3） was found to bind to W-box by screening a cDNA library of T. androssowii with yeast one-hybrid system. TaMCM3 failed in binding to W-box, when ＂G＂ mutated to ＂A＂ or ＂A＂ mutated to ＂G＂ of core ＂TGAC＂ sequence. Therefore, the bind- ing of TaMCM3 to W-box is specific. Moreover, the TaMCM3 is binding to the promoter fragment containing W-box, but the binding is loosed when W-box is mutated. TaMCM3 interacted with promoter of TaelFSA1 through binding to W-box. TaMCM3 can specifically recognize W-box and be served as transcription factor.

关 键 词：柽柳 EIF5A 启动子 W—box 酵母单杂交 

分 类 号：Q943.2[生物学—植物学]