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Molecular Mechanism of Regulation of Cell Cycle-Dependent Gene Expression in Higher Plants

高等植物细胞周期依赖性基因表达调控的分子机制

基本信息

批准号：
01440002
负责人：
IWABUCHI Masaki
金额：
$ 20.48万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01440002/
关键词：
Wheat histone genes S-phase-specific transcription Cis-acting elements Trans-acting factors DNA-binding protein bZip structure Zn-finger motif Autoregulation 転写調節機構 HBPー1 HBPー2 Zn・フィンガ-モチ-フコムギ・ヒストンH3遺伝子 S期特異的発現シス制御エレメント DNA

项目摘要

In order to elucidate the molecular mechanisms of S-phase-specific transcription regulation in the division cycle of higher plant cells, my co-workers and I have studied the structures and functions of cis-acting elements and trans-acting factors which are implicated in the regulation of the wheat histone H3 gene transcription. We have obtained the following results in this research.1. As cis-elements for the S-phase specificity of the H3 gene transcription we have identified two motifs (hexamer and octamer sequences) in three cis-elements located within the -185 region of the H3 promoter.2. We have identified the sequence-specific DNA-binding proteins, HBP-1a and HBP-1b, that are interact with the hexamer motif. The CDNA clones encoding each of proteins were isolated and their nucleotide sequences were determined. The amino acid sequences deduced from the nucleotide sequences suggested that the HBP-la and -1b have a bZIP motif at the regions near the amino- and carboxy-termini, respectively. We also have identified a DNA-binding protein, HBP-2, which specifically interacts with another cis-element, nonamer sequence, and this protein was shown to have a Zn-finger motif.3. To study the function of HBP-1a and -1b, we examined the influence of the effector gene consisting of the CaMV 35S promoter and HBP-1a or -1b EDNA on the expression of the reporter gene containing the H3 promoter/GUS coding region in the transient expression system using rice and tabacco cell cultures. We have obtained evidence that the proteins both are regulatory factors involved in the H3 gene transcription.4. We found the hexamer and hexamer-like motifs in the promoter regions of cloned HBP-1a and -1b genes and observed that the motifs can bind HBP-1a and -1b. It is suggested that transcription of these genes may be autoregulated by the factors themselves.

为了阐明高等植物细胞分裂周期中S期特异性转录调控的分子机制，我和我的同事研究了小麦组蛋白H3基因转录调控中涉及的顺式作用元件和反式作用因子的结构和功能。本研究取得了以下成果：1.作为H3基因转录S期特异性的顺式元件，我们在位于H3启动子-185区域的三个顺式元件中鉴定了两个基序（六聚体和八聚体序列）.我们已经确定了序列特异性DNA结合蛋白，HBP-1a和HBP-1b，与六聚体基序相互作用。分离编码每种蛋白质的cDNA克隆，并测定其核苷酸序列。从核苷酸序列推导的氨基酸序列表明，HBP-la和-1b分别在氨基和羧基末端附近的区域具有bZIP基序。我们还鉴定了一个DNA结合蛋白HBP-2，它与另一个顺式元件九聚体序列特异性相互作用，并且该蛋白具有锌指基序.为了研究HBP-1a和-1b的功能，我们用水稻和烟草细胞培养物研究了由CaMV 35 S启动子和HBP-1a或-1b EDNA组成的效应基因对含有H3启动子/GUS编码区的报告基因在瞬时表达系统中表达的影响。我们获得的证据表明这两种蛋白都是参与H3基因转录的调控因子.我们在克隆的HBP-1a和-1b基因的启动子区发现了六聚体和六聚体样基序，并观察到这些基序可以与HBP-1a和-1b结合。这表明，这些基因的转录可能是自动调节的因素本身。