A Membrane-Bound NAC Transcription Factor, ANAC017, Mediates Mitochondrial Retrograde Signaling in Arabidopsis
A Membrane-Bound NAC Transcription Factor, ANAC017, Mediates Mitochondrial Retrograde Signaling in Arabidopsis
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DOI：
10.1105/tpc.113.113985
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发表时间：
2013-09-01
期刊：
影响因子：
11.6
通讯作者：
Giraud, Estelle
中科院分区：
文献类型：
--
作者：
Ng, Sophia;Ivanova, Aneta;Giraud, Estelle
植物需要每日协调调节能量代谢以获得最佳生长和存活，因此需要将细胞反应与线粒体和质体逆行信号整合。使用正向遗传筛选来表征交替氧化酶1a（rao）突变体的调节剂，我们鉴定RAO 2/拟南芥NAC结构域包含蛋白17（ANAC 017）作为AOX 1a的直接正调节剂。RAO 2/ANAC 017通过C-末端跨膜（TM）结构域靶向内质网（ER）和F-肌动蛋白中的连接和连接。一个共有的菱形蛋白酶切割位点存在于ANAC 017中，就在预测的TM结构域之前。此外，添加菱形蛋白酶抑制剂N-p-甲苯磺酰基-L-Phe氯甲基消除了抗霉素A处理后AOX 1a的诱导。用N-末端红色荧光蛋白（RFP）和C-末端绿色荧光蛋白（GFP）同时荧光标记ANAC 017揭示了N-末端RFP结构域迁移到细胞核中，而C-末端GFP标签保留在ER中。在胁迫处理下由RAO 2/ANAC 017调节的转录网络的全基因组分析揭示，RAO 2/ANAC 017功能对于作为对胞质过氧化氢（H2 O2）的主要响应而观察到的> 85%的变化是必需的，但仅对于响应于抗霉素A处理而观察到的33%的转录变化是必需的。具有突变的rao 2/anac 017的植物对胁迫更敏感，而功能获得性突变导致植物在未处理的条件下具有较低的H2 O2细胞水平。
Plants require daily coordinated regulation of energy metabolism for optimal growth and survival and therefore need to integrate cellular responses with both mitochondrial and plastid retrograde signaling. Using a forward genetic screen to characterize regulators of alternative oxidase1a (rao) mutants, we identified RAO2/Arabidopsis NAC domain-containing protein17 (ANAC017) as a direct positive regulator of AOX1a. RAO2/ANAC017 is targeted to connections and junctions in the endoplasmic reticulum (ER) and F-actin via a C-terminal transmembrane (TM) domain. A consensus rhomboid protease cleavage site is present in ANAC017 just prior to the predicted TM domain. Furthermore, addition of the rhomboid protease inhibitor N-p-Tosyl-L-Phe chloromethyl abolishes the induction of AOX1a upon antimycin A treatment. Simultaneous fluorescent tagging of ANAC017 with N-terminal red fluorescent protein (RFP) and C-terminal green fluorescent protein (GFP) revealed that the N-terminal RFP domain migrated into the nucleus, while the C-terminal GFP tag remained in the ER. Genome-wide analysis of the transcriptional network regulated by RAO2/ANAC017 under stress treatment revealed that RAO2/ANAC017 function was necessary for > 85% of the changes observed as a primary response to cytosolic hydrogen peroxide (H2O2), but only; 33% of transcriptional changes observed in response to antimycin A treatment. Plants with mutated rao2/anac017 were more stress sensitive, whereas a gain-of-function mutation resulted in plants that had lower cellular levels of H2O2 under untreated conditions.