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Functional analysis of a rice importin-α binding protein

水稻导入蛋白-α结合蛋白的功能分析

基本信息

批准号：
13640643
负责人：
IWASAKI Toshisuke
金额：
$ 2.3万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13640643/
关键词：
Nuclear import of proteins Response to light environment Protein-protein interactions TPR motif Rice(Oryza sativa)Arabidopsis thaliana T-DNA tag line Light-responsive gene expression 核タンパク質インポーティンα結合タンパク質核局在化シグナル

项目摘要

To identify novel nuclear proteins that are involved in plants' responses to light environment, the author analyzed the function of a candidate protein, named IABP4, that had been isolated by its binding to rice importin-α1a(IMPα1a), a component of nuclear transport machinery, whose expression is strong in the dark and is down-regulated by light. The obtained results are as follows.1.Gene expression and the effect of light : Similar to rice IMPα1a gene, the expression of IABP4 was down-regulated by light both in etiolated-and green-seedlings of rice and Arabidopsis. The result suggests that nuclear accumulation of IABP4 might be reduced in the dark as a result of reduced IABP4 protein level as well as reduced nuclear import of IAB P4.2.Functional analysis by knockout lines : Five knockout lines of IABP4 were identified in Arabidopsis, and the proportion of seedlings with open hook and cotyledons in the dark was higher in the knockouts than in wild type, suggesting that IABP4 protein may be suppressing photomorphogenesis in the dank. Because germinated seedlings were all hetero-zygous with respect to T-DNA insertion, the homozygote is considered to be embryonic lethal or unable to germinate, suggesting that IABP4 is not only involved in the control of photomorphgenesis but also essential for plants.3.IntraceMar localization and the effect of light: Though we obtained IABP4 antiserum, it contained antibodies to other plant proteins, so we could not use it for localization studies. By purifying antibody or by using GFP-fusion, examination of nuclear localization of IABP4 is urgently required.

为了鉴定参与植物对光环境反应的新型核蛋白，作者分析了一种名为IABP 4的候选蛋白的功能，该蛋白是通过与水稻输入蛋白-α1a（IMPα1a）结合而分离出来的。核运输机制的组成部分，其表达在黑暗中很强，并被光下调。主要研究结果如下：1.基因表达及光的影响：与水稻IMPα1a基因相似，水稻和拟南芥黄化苗和绿苗中IABP 4的表达均受光的下调。结果表明，由于IABP 4蛋白水平降低以及IABP 4的核输入减少，IABP 4的核积累在黑暗中可能减少。在拟南芥中鉴定了5个IABP 4基因敲除株系，敲除株系中具有开钩和子叶的幼苗比例高于野生型，这表明IABP 4蛋白可能抑制潮湿中的光形态建成。由于萌发的幼苗在T-DNA插入方面都是杂合的，因此纯合子被认为是胚胎致死的或不能萌发的，这表明IABP 4不仅参与光形态建成的控制，而且对植物是必不可少的。虽然我们获得了IABP 4抗血清，但它含有其他植物蛋白的抗体，因此我们不能将其用于定位研究。通过纯化抗体或利用GFP融合技术，对IABP 4的核定位进行检测是迫切需要的。