Yang Xiaoqing, Lee Sungsu, So Jai-Hyun, Dharmasiri Suni, Dharmasiri Nihal, Ge Lei, Jensen Carolyn, Hangarter Roger, Hobbie Lawrence, Estelle Mark
Department of Biology, Indiana University, Bloomington, IN 47405, USA.
Plant J. 2004 Dec;40(5):772-82. doi: 10.1111/j.1365-313X.2004.02254.x.
Recent studies of auxin response have focused on the functions of three sets of proteins: the auxin (Aux) response factors (ARFs), the Aux/IAAs, and the F-box protein TIR1. The ARF proteins bind DNA and directly activate or repress transcription of target genes while the Aux/IAA proteins repress ARF function. TIR1 is part of a ubiquitin protein ligase required for degradation of Aux/IAA proteins. Here we report the isolation and characterization of a novel mutant of Arabidopsis called axr5-1. Mutant plants are resistant to auxin and display a variety of auxin-related growth defects including defects in root and shoot tropisms. Further, the axr5-1 mutation results in a decrease in auxin-regulated transcription. The molecular cloning of AXR5 revealed that the gene encodes the IAA1 protein, a member of the Aux/IAA family of proteins. AXR5 is expressed throughout plant development consistent with the pleiotropic mutant phenotype. The axr5-1 mutation results in an amino acid substitution in conserved domain II of the protein, similar to gain-of-function mutations recovered in other members of this gene family. Biochemical studies show that IAA1/AXR5 interacts with TIR1 in an auxin-dependent manner. The mutation prevents this interaction suggesting that the mutant phenotype is caused by the accumulation of IAA1/AXR5. Our results provide further support for a model in which most members of the Aux/IAA family are targeted for degradation by SCFTIR1 in response to auxin.
生长素(Aux)应答因子(ARF)、Aux/IAAs和F-box蛋白TIR1。ARF蛋白结合DNA并直接激活或抑制靶基因的转录,而Aux/IAA蛋白则抑制ARF的功能。TIR1是Aux/IAA蛋白降解所需的泛素蛋白连接酶的一部分。在此,我们报告了拟南芥一种名为axr5-1的新突变体的分离和鉴定。突变植株对生长素具有抗性,并表现出多种与生长素相关的生长缺陷,包括根和茎向性的缺陷。此外,axr5-1突变导致生长素调节的转录减少。AXR5的分子克隆表明该基因编码IAA1蛋白,它是Aux/IAA蛋白家族的成员之一。AXR5在植物发育过程中均有表达,这与多效性突变体表型一致。axr5-1突变导致该蛋白保守结构域II中的一个氨基酸替换,类似于在该基因家族其他成员中发现的功能获得性突变。生化研究表明,IAA1/AXR5以生长素依赖的方式与TIR1相互作用。该突变阻止了这种相互作用,表明突变体表型是由IAA1/AXR5的积累引起的。我们的结果为一个模型提供了进一步的支持,即Aux/IAA家族的大多数成员在生长素的作用下被SCFTIR1靶向降解。