Terada R, Nakayama T, Iwabuchi M, Shimamoto K
Plantech Research Institute, Yokohama, Japan.
Plant Mol Biol. 1995 Jan;27(1):17-26. doi: 10.1007/BF00019175.
Type I element (CCACGTCACCGATCCGCG) is a well-conserved regulatory element found in proximal promoter region of a certain class of plant histone genes, that is composed of two independent cis-acting elements of the hexamer (ACGTCA) and the reverse-oriented octamer (GATCCGCG) motifs. To investigate functional role(s) of the type I element in regulation of a wheat histone H3 gene (TH012) promoter activity in vivo, base substitution mutations were introduced into the element and activities of the mutated promoters were examined in cultured rice cells, and in regenerated roots and anther walls of transgenic rice plants by employing a GUS reporter system. Mutations of each or both of the hexamer and the octamer motifs caused a reduction in the promoter activity in protoplasts transfected transiently or stably transformed calli. The mutation of the octamer motif with or without the mutation of the hexamer motif caused a marked reduction of the promoter activity in the root meristem of transgenic rice although the mutation of the hexamer motif alone caused a weak reduction. In contrast to these results, no effect of the mutations of either the hexamer or the octamer motif was found in the anther wall in which replication-independent activity of the H3 promoter was observed. Our results suggested that the hexamer and the octamer motifs may play important role(s) in regulation of replication-dependent but not of replication-independent expression of the wheat histone H3 gene.
I型元件(CCACGTCACCGATCCGCG)是在某类植物组蛋白基因的近端启动子区域发现的一个高度保守的调控元件,它由六聚体(ACGTCA)和反向八聚体(GATCCGCG)基序的两个独立顺式作用元件组成。为了研究I型元件在体内调控小麦组蛋白H3基因(TH012)启动子活性中的功能作用,将碱基替代突变引入该元件,并通过使用GUS报告系统在培养的水稻细胞以及转基因水稻植株的再生根和花药壁中检测突变启动子的活性。六聚体和八聚体基序各自或两者的突变导致瞬时转染的原生质体或稳定转化的愈伤组织中启动子活性降低。八聚体基序的突变无论六聚体基序是否突变,都会导致转基因水稻根分生组织中启动子活性显著降低,尽管仅六聚体基序的突变导致的降低较弱。与这些结果相反,在观察到H3启动子非复制依赖性活性的花药壁中,未发现六聚体或八聚体基序的突变有任何影响。我们的结果表明,六聚体和八聚体基序可能在调控小麦组蛋白H3基因的复制依赖性而非非复制依赖性表达中起重要作用。