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拟南芥细胞色素c氧化酶亚基5b的一种同工型编码基因COX5b-2中参与蔗糖和UV-B光诱导表达的调控元件的鉴定。

Identification of regulatory elements involved in expression and induction by sucrose and UV-B light of the Arabidopsis thaliana COX5b-2 gene, encoding an isoform of cytochrome c oxidase subunit 5b.

作者信息

Comelli Raúl N, Gonzalez Daniel H

机构信息

Instituto de Agrobiotecnología del Litoral (CONICET-UNL), Cátedra de Biología Celular y Molecular, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, CC 242 Paraje El Pozo, Santa Fe, Argentina.

出版信息

Physiol Plant. 2009 Nov;137(3):213-24. doi: 10.1111/j.1399-3054.2009.01285.x. Epub 2009 Aug 26.

Abstract

The promoter sequences required for expression of the Arabidopsis thaliana COX5b-2 gene, encoding an isoform of cytochrome c oxidase subunit 5b, were analyzed using plants transformed with deleted and mutagenized forms of the promoter fused to gus. A 1000-bp promoter fragment produces expression in root and shoot meristems, leaf and cotyledon tips, and anthers. Deletion analysis indicated the presence of positive and negative regulatory elements. A regulatory element located between -660 and -620 from the translation start site was identified as a G-box by mutagenic analysis. Mutation of the G-box, that is present within the coding region of the preceding gene in the genome, increases expression of COX5b-2 in cotyledon and leaf lamina and abolishes induction by ultraviolet-B (UV-B) light, which presumably acts through the removal of an inhibitory factor. Identified positive regulatory elements include a site II element (TGGGCC), a related element with the sequence TGGGTC and four initiator elements (YTCANTYY) that completely abolish expression when mutated in combination. Site II elements are also involved in the response to sucrose. The results imply that the COX5b-2 gene has retained expression characteristics presented by most respiratory chain component genes, but its expression mechanisms have diverged from those employed by COX5b-1, the other gene encoding cytochrome c oxidase subunit 5b in Arabidopsis.

摘要

利用与gus融合的缺失和诱变形式的启动子转化的植物,分析了编码细胞色素c氧化酶亚基5b同工型的拟南芥COX5b-2基因表达所需的启动子序列。一个1000bp的启动子片段在根和茎分生组织、叶和子叶尖端以及花药中产生表达。缺失分析表明存在正调控元件和负调控元件。通过诱变分析,确定位于翻译起始位点-660至-620之间的调控元件为G-盒。基因组中前一个基因编码区域内存在的G-盒发生突变,会增加COX5b-2在子叶和叶片中的表达,并消除紫外线-B(UV-B)光的诱导作用,推测这是通过去除一种抑制因子来实现的。已确定的正调控元件包括一个II型元件(TGGGCC)、一个序列为TGGGTC的相关元件和四个起始元件(YTCANTYY),当它们组合突变时会完全消除表达。II型元件也参与对蔗糖的反应。结果表明,COX5b-2基因保留了大多数呼吸链组分基因呈现的表达特征,但其表达机制与拟南芥中另一个编码细胞色素c氧化酶亚基5b的基因COX5b-1所采用的机制有所不同。

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