Department of Energy-Plant Research Laboratory, Michigan State University, East Lansing, MI 48824, USA.
Plant J. 2010 Aug;63(4):613-22. doi: 10.1111/j.1365-313X.2010.04265.x.
Jasmonates (JAs) are fatty acid-derived signaling compounds that control diverse aspects of plant growth, development and immunity. The F-box protein COI1 functions both as a receptor for jasmonoyl-l-isoleucine (JA-Ile) and as the component of an E3-ubiquitin ligase complex (SCF(COI1) ) that targets JAZ transcriptional regulators for degradation. A key feature of JAZ proteins is the C-terminal Jas motif that mediates the JA-Ile-dependent interaction with COI1. Here, we show that most JAZ genes from evolutionarily diverse plants contain a conserved intron that splits the Jas motif into 20 N-terminal and seven C-terminal (X(5) PY) amino acid submotifs. In most members of the Arabidopsis JAZ family, alternative splicing events involving retention of this intron generate proteins that are truncated before the X(5) PY sequence. In vitro pull-down and yeast two-hybrid assays indicate that these splice variants have reduced capacity to form stable complexes with COI1 in the presence of the bioactive stereoisomer of the hormone (3R,7S)-JA-Ile. cDNA overexpression studies showed that some, but not all, truncated splice variants are dominant repressors of JA signaling. We also show that strong constitutive expression of an intron-containing JAZ10 genomic clone is sufficient to repress JA responses. These findings provide evidence for functional differences between JAZ isoforms, and establish a direct link between the alternative splicing of JAZ pre-mRNA and the dominant repression of JA signal output. We propose that production of dominant JAZ repressors by alternative splicing reduces the negative consequences associated with inappropriate or hyperactivation of the JA response pathway.
茉莉酸(JAs)是脂肪酸衍生的信号化合物,可控制植物生长、发育和免疫的多个方面。F-box 蛋白 COI1 既是茉莉酰基异亮氨酸(JA-Ile)的受体,又是 E3-泛素连接酶复合物(SCF(COI1))的组成部分,该复合物可将 JAZ 转录调节剂靶向降解。JAZ 蛋白的一个关键特征是 C 端的 Jas 基序,该基序介导 JA-Ile 依赖的与 COI1 的相互作用。在这里,我们表明来自进化上不同植物的大多数 JAZ 基因都含有一个保守的内含子,该内含子将 Jas 基序分裂成 20 个 N 端和七个 C 端(X(5)PY)氨基酸亚基序。在拟南芥 JAZ 家族的大多数成员中,涉及保留该内含子的剪接事件会产生在 X(5)PY 序列之前截断的蛋白质。体外下拉和酵母双杂交测定表明,在激素(3R,7S)-JA-Ile 的生物活性立体异构体存在的情况下,这些剪接变体与 COI1 形成稳定复合物的能力降低。cDNA 过表达研究表明,一些但不是所有的截短剪接变体都是 JA 信号的显性抑制剂。我们还表明,包含内含子的 JAZ10 基因组克隆的强组成型表达足以抑制 JA 反应。这些发现为 JAZ 同工型之间存在功能差异提供了证据,并建立了 JAZ 前体 mRNA 选择性剪接与 JA 信号输出的显性抑制之间的直接联系。我们提出,通过选择性剪接产生显性 JAZ 抑制剂可降低与 JA 反应途径的不当或过度激活相关的负面后果。