Boter Marta, Ruíz-Rivero Omar, Abdeen Ashraf, Prat Salomé
Departament de Genètica Molecular, Institut de Biologia Molecular de Barcelona, CID-CSIC, 08034 Barcelona, Spain.
Genes Dev. 2004 Jul 1;18(13):1577-91. doi: 10.1101/gad.297704.
Jasmonates (JA) are important regulators of plant defense responses that activate expression of many wound-induced genes including the tomato proteinase inhibitor II (pin2) and leucine aminopeptidase (LAP) genes. Elements required for JA induction of the LAP gene are all present in the -317 to -78 proximal promoter region. Using yeast one-hybrid screening, we have identified the bHLH-leu zipper JAMYC2 and JAMYC10 proteins, specifically recognizing a T/G-box AACGTG motif in this promoter fragment. Mutation of the G-box element decreases JA-responsive LAP promoter expression. Expression of JAMYC2 and JAMYC10 is induced by JA, with a kinetics that precedes that of the LAP or pin2 transcripts. JAMYC overexpression enhanced JA-induced expression of these defense genes in potato, but did not result in constitutive transcript accumulation. Using footprinting assays, an additional protected element was identified, located directly adjacent to the T/G-box motif. Mutation of this element abolishes JA response, showing that recognition of this duplicated element is also required for gene expression. Knockout mutants in the AtMYC2 homolog gene of Arabidopsis are insensitive to JA and exhibit a decreased activation of the JA-responsive genes AtVSP and JR1. Activation of the PDF1.2 and b-CHI, ethylene/JA-responsive genes, is, however, increased in these mutants. These results show that the JAMYC/AtMYC2 transcription factors function as members of a MYC-based regulatory system conserved in dicotyledonous plants with a key role in JA-induced defense gene activation.
茉莉酸(JA)是植物防御反应的重要调节因子,可激活许多创伤诱导基因的表达,包括番茄蛋白酶抑制剂II(pin2)和亮氨酸氨肽酶(LAP)基因。JA诱导LAP基因所需的元件都存在于-317至-78近端启动子区域。通过酵母单杂交筛选,我们鉴定出了bHLH - 亮氨酸拉链JAMYC2和JAMYC10蛋白,它们特异性识别该启动子片段中的T/G - 盒AACGTG基序。G - 盒元件的突变会降低JA响应的LAP启动子表达。JAMYC2和JAMYC10的表达由JA诱导,其动力学先于LAP或pin2转录本。JAMYC过表达增强了JA诱导的这些防御基因在马铃薯中的表达,但并未导致组成型转录本积累。通过足迹分析,鉴定出了一个额外的受保护元件,它直接毗邻T/G - 盒基序。该元件的突变消除了JA响应,表明对这个重复元件的识别也是基因表达所必需的。拟南芥AtMYC2同源基因的敲除突变体对JA不敏感,并且JA响应基因AtVSP和JR1的激活降低。然而,乙烯/JA响应基因PDF1.2和b - CHI在这些突变体中的激活增加。这些结果表明,JAMYC/AtMYC2转录因子作为双子叶植物中保守的基于MYC的调控系统的成员发挥作用,在JA诱导的防御基因激活中起关键作用。