拟南芥rd29A基因在脱水和高盐胁迫响应中ABA依赖型表达过程中两个顺式作用元件ABRE和DRE之间的相互作用

Interaction between two cis-acting elements, ABRE and DRE, in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses.

作者信息

Narusaka Yoshihiro, Nakashima Kazuo, Shinwari Zabta K, Sakuma Yoh, Furihata Takashi, Abe Hiroshi, Narusaka Mari, Shinozaki Kazuo, Yamaguchi-Shinozaki Kazuko

机构信息

Biological Resources Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 305-8686, Japan.

出版信息

Plant J. 2003 Apr;34(2):137-48. doi: 10.1046/j.1365-313x.2003.01708.x.

DOI:10.1046/j.1365-313x.2003.01708.x

Abstract

Many abiotic stress-inducible genes contain two cis-acting elements, namely a dehydration-responsive element (DRE; TACCGACAT) and an ABA-responsive element (ABRE; ACGTGG/TC), in their promoter regions. We precisely analyzed the 120 bp promoter region (-174 to -55) of the Arabidopsis rd29A gene whose expression is induced by dehydration, high-salinity, low-temperature, and abscisic acid (ABA) treatments and whose 120 bp promoter region contains the DRE, DRE/CRT-core motif (A/GCCGAC), and ABRE sequences. Deletion and base substitution analyses of this region showed that the DRE-core motif functions as DRE and that the DRE/DRE-core motif could be a coupling element of ABRE. Gel mobility shift assays revealed that DRE-binding proteins (DREB1s/CBFs and DREB2s) bind to both DRE and the DRE-core motif and that ABRE-binding proteins (AREBs/ABFs) bind to ABRE in the 120 bp promoter region. In addition, transactivation experiments using Arabidopsis leaf protoplasts showed that DREBs and AREBs cumulatively transactivate the expression of a GUS reporter gene fused to the 120 bp promoter region of rd29A. These results indicate that DRE and ABRE are interdependent in the ABA-responsive expression of the rd29A gene in response to ABA in Arabidopsis.

摘要

许多非生物胁迫诱导基因在其启动子区域含有两个顺式作用元件，即脱水响应元件（DRE；TACCGACAT）和脱落酸响应元件（ABRE；ACGTGG/TC）。我们精确分析了拟南芥rd29A基因120bp的启动子区域（-174至-55），该基因的表达受脱水、高盐、低温和脱落酸（ABA）处理诱导，其120bp启动子区域包含DRE、DRE/CRT核心基序（A/GCCGAC）和ABRE序列。对该区域的缺失和碱基替换分析表明，DRE核心基序作为DRE发挥作用，且DRE/DRE核心基序可能是ABRE的偶联元件。凝胶迁移率变动分析显示，DRE结合蛋白（DREB1s/CBFs和DREB2s）与DRE和DRE核心基序均结合，而ABRE结合蛋白（AREBs/ABFs）与120bp启动子区域中的ABRE结合。此外，使用拟南芥叶原生质体的反式激活实验表明，DREBs和AREBs可累积反式激活与rd29A基因120bp启动子区域融合的GUS报告基因的表达。这些结果表明，在拟南芥中，DRE和ABRE在rd29A基因响应ABA的ABA应答表达中相互依赖。