Zhang Botao, Van Aken Olivier, Thatcher Louise, De Clercq Inge, Duncan Owen, Law Simon R, Murcha Monika W, van der Merwe Margaretha, Seifi Hamed Soren, Carrie Chris, Cazzonelli Christopher, Radomiljac Jordan, Höfte Monica, Singh Karam B, Van Breusegem Frank, Whelan James
ARC Centre of Excellence in Plant Energy Biology, University of Western Australia, Bayliss Building M316, 35 Stirling Highway, Crawley, WA, 6009, Australia; Department of Botany, ARC Centre of Excellence in Plant Energy Biology, School of Life Science, La Trobe University, Bundoora, Vic., 3086, Australia.
Plant J. 2014 Nov;80(4):709-27. doi: 10.1111/tpj.12665.
One of the most stress-responsive genes encoding a mitochondrial protein in Arabidopsis (At3g50930) has been annotated as AtBCS1 (cytochrome bc1 synthase 1), but was previously functionally uncharacterised. Here, we show that the protein encoded by At3g50930 is present as a homo-multimeric protein complex on the outer mitochondrial membrane and lacks the BCS1 domain present in yeast and mammalian BCS1 proteins, with the sequence similarity restricted to the AAA ATPase domain. Thus we propose to re-annotate this protein as AtOM66 (Outer Mitochondrial membrane protein of 66 kDa). While transgenic plants with reduced AtOM66 expression appear to be phenotypically normal, AtOM66 over-expression lines have a distinct phenotype, showing strong leaf curling and reduced starch content. Analysis of mitochondrial protein content demonstrated no detectable changes in mitochondrial respiratory complex protein abundance. Consistent with the stress inducible expression pattern, over-expression lines of AtOM66 are more tolerant to drought stress but undergo stress-induced senescence earlier than wild type. Genome-wide expression analysis revealed a constitutive induction of salicylic acid-related (SA) pathogen defence and cell death genes in over-expression lines. Conversely, expression of SA marker gene PR-1 was reduced in atom66 plants, while jasmonic acid response genes PDF1.2 and VSP2 have increased transcript abundance. In agreement with the expression profile, AtOM66 over-expression plants show increased SA content, accelerated cell death rates and are more tolerant to the biotrophic pathogen Pseudomonas syringae, but more susceptible to the necrotrophic fungus Botrytis cinerea. In conclusion, our results demonstrate a role for AtOM66 in cell death and amplifying SA signalling.
拟南芥中编码一种线粒体蛋白的应激反应最强的基因之一(At3g50930)已被注释为AtBCS1(细胞色素bc1合酶1),但此前其功能尚未得到表征。在这里,我们表明,At3g50930编码的蛋白质以同多聚体蛋白复合物的形式存在于线粒体外膜上,并且缺乏酵母和哺乳动物BCS1蛋白中存在的BCS1结构域,序列相似性仅限于AAA ATP酶结构域。因此,我们建议将该蛋白重新注释为AtOM66(66 kDa线粒体外膜蛋白)。虽然AtOM66表达降低的转基因植物在表型上似乎正常,但AtOM66过表达株系具有明显的表型,表现出强烈的叶片卷曲和淀粉含量降低。线粒体蛋白含量分析表明,线粒体呼吸复合物蛋白丰度没有可检测到的变化。与应激诱导的表达模式一致,AtOM66过表达株系对干旱胁迫更耐受,但比野生型更早经历应激诱导的衰老。全基因组表达分析揭示了过表达株系中水杨酸相关(SA)病原体防御和细胞死亡基因的组成型诱导。相反,atom66植物中SA标记基因PR-1的表达降低,而茉莉酸反应基因PDF1.2和VSP2的转录本丰度增加。与表达谱一致,AtOM66过表达植物显示SA含量增加、细胞死亡率加快,对活体营养型病原体丁香假单胞菌更耐受,但对坏死营养型真菌灰葡萄孢更敏感。总之,我们的结果证明了AtOM66在细胞死亡和放大SA信号传导中的作用。