Analysis of interaction between transcription factors in the regulation of wheat histone gene expression

转录因子在小麦组蛋白基因表达调控中的相互作用分析

• 批准号: 05640729
• 负责人: MESHI Tetsuo
• 金额: $ 1.28万
• 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 (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05640729/
• 关键词: histone gene; interaction; transcription regulation; bZIP-type factor; transcription factor; HBP-1a

To identify the components involved in the formation of the special complex on the type I element in the wheat histone H3 promoter, which is known to confer the S-phase specific transcription, we carried out West-western screening with phosphorylated HBP-1a (17) as a probe. Aa a result, a novel bZIP-type transcription factor, HALF-1, was isolated from wheat cDNA library. With GST-fusions and by gel shift analysis, HALF-1 and HBP-1a (17) were demonstrated to interact with each other through their ZIP domains. Northern analysis revealed that HALF-1 expressed highly in the late S to early G2 phases. Transient assay revealed that HALF-1 had a potential activation domain that contained an amino acid stretch conserved among several plant bZIP proteins. The DNA-binding specificity and the structural characteristics showed that HALF-1 belonged to the HBP-1a subfamily.Wheat nuclear extracts prepared from S phase cells contained some specific DNA-binding activity directing to the hexamer motif and to the octamer motif. The S-phase specific hexamer-binding activity had the character similar to that of the HBP-1a subfamily and was suggested to be regulated by phosphorylation.

为了确定小麦组蛋白H3启动子中I型元件上的特殊复合体的形成所涉及的成分，我们以磷酸化的HBP-1a(17)为探针进行了West-Western筛选。结果从小麦cDNA文库中克隆到一个新的bZIP型转录因子Half-1。GST融合和凝胶位移分析表明，Half-1和HBP-1a(17)通过ZIP结构域相互作用。Northern分析表明，Half-1在S晚期至G2期早期高表达。瞬时分析表明，Half-1具有潜在的激活结构域，含有一个在几种植物bZIP蛋白中保守的氨基酸延伸。DNA结合的特异性和结构特征表明，Half-1属于HBP-1a亚家族。从S期细胞中提取的小麦核提取物具有针对六聚体基序和八聚体基序的特异性DNA结合活性。S时相特异性六聚体结合活性与HBP-1a亚家族相似，被认为是通过磷酸化来调节的。

项目成果

Ohtsubo,N.: "The conserved 3'-flanking sequence.AATGGAAATG,of the wheat histone H3 gene is necessary for the accurate 3'-end formation of mRNA." Nucl.Acids Res.22. 1052-1058 (1994)

Ohtsubo,N.：“小麦组蛋白 H3 基因的保守 3 侧翼序列 AATGGAAATG 对于 mRNA 的准确 3 端形成是必需的。”

Minami,M: "Coordinate gene expression of five subclass histones and the putative transcription factors,HBP-la and HBP-lb,of histone genes in wheat." PlantMol.Biol,. 23. 429-434 (1993)

Minami,M：“协调小麦中五个组蛋白亚类和假定的组蛋白基因转录因子 HBP-la 和 HBP-lb 的基因表达。”

Huh,G.H: "Differential expression of the twe types of histone H2A genes in wheat." Biochim.Biophys.Acta.1261. 155-160 (1995)

Huh，G.H：“两种类型的组蛋白 H2A 基因在小麦中的差异表达。”

Terada,R: "A wheat histone H3 promoter confers cell division-dependent and-independent expression of the gusA gene in transgenic rice plants." PlantJ.2. 1049-1056 (1993)

Terada,R：“小麦组蛋白 H3 启动子在转基因水稻植物中赋予 gusA 基因细胞分裂依赖性和独立性表达。”

Huh,G.H.: "Differential expression of the two types of histone H2A genes in wheat." Biochim.Biophys.Acta.(印刷中). (1995)

Huh，G.H.：“小麦中两种类型的组蛋白 H2A 基因的差异表达。”（正在出版）。