Selective autophagy regulates heat stress memory in Arabidopsis by NBR1-mediated targeting of HSP90.1 and ROF1.
Selective autophagy regulates heat stress memory in Arabidopsis by NBR1-mediated targeting of HSP90.1 and ROF1.
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选择性自噬通过NBR1介导的HSP90.1和ROF1靶向调控拟南芥的热应激记忆。
DOI：
10.1080/15548627.2020.1820778
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发表时间：
2021-09
期刊：
影响因子：
13.3
通讯作者：
Balazadeh S
中科院分区：
文献类型：
--
作者：
Thirumalaikumar VP;Gorka M;Schulz K;Masclaux-Daubresse C;Sampathkumar A;Skirycz A;Vierstra RD;Balazadeh S
在自然界中，植物不断地暴露于许多短暂的，但反复出现的压力。因此，为了完成它们的生命周期，植物需要在胁迫停止后恢复的能力和维持胁迫记忆之间保持动态平衡。最近，我们发现了一个新的功能作用的macroautophagy/自噬调节恢复热应激（HS）和重置细胞记忆的HS在拟南芥。在这里，我们证明了NBR 1（BRCA 1基因1旁边）在HS恢复过程中作为选择性自噬的受体发挥着至关重要的作用。免疫印迹分析和共聚焦显微镜显示，水平的NBR 1蛋白，NBR 1标记的斑点，和NBR 1的活性都高于在HS恢复阶段比以前。与NBR 1相互作用的蛋白质的免疫共沉淀分析和NBR 1无效突变体和野生型植物的比较蛋白质组学分析确定了58种蛋白质作为NBR 1的潜在新靶点。细胞、生物化学和功能遗传学研究证实，NBR 1与热休克蛋白90.1（heat shock protein 90.1，HSP）和FKBP家族成员ROF 1（rotamase FKBP 1，ROF 1）相互作用，并通过自噬介导它们的降解，自噬通过减弱由HSFA 2转录因子调节的HSP基因的表达来抑制对HS的反应。因此，NBR 1的功能丧失突变导致更强的HS记忆表型。总之，我们的研究结果提供了新的见解的机械原理，自噬调节植物对复发性HS的反应。缩略语：目的：Atg 8相互作用基序; ATG：自噬相关; BiFC：双分子荧光互补; ConA：伴卡那霉素A; CoIP：免疫共沉淀; DMSO：二甲亚砜; FKBP：FK 506结合蛋白; FBPASE：果糖1，6-二磷酸酶; GFP：绿色荧光蛋白; HS：热应激; HSF：热休克因子; HSFA 2：热休克因子A2; HSP：热休克蛋白; HSP 90：热休克蛋白90; LC-MS/MS：液相色谱-串联质谱; 3-MA：3-甲基腺嘌呤; NBR 1：BRCA 1旁; PQC：蛋白质质量控制; RFP：红色荧光蛋白; ROF1：旋转异构酶FKBP 1; TF：转录因子;浴缸：微管蛋白;乌巴：泛素相关; YFP：黄色荧光蛋白
In nature, plants are constantly exposed to many transient, but recurring, stresses. Thus, to complete their life cycles, plants require a dynamic balance between capacities to recover following cessation of stress and maintenance of stress memory. Recently, we uncovered a new functional role for macroautophagy/autophagy in regulating recovery from heat stress (HS) and resetting cellular memory of HS in Arabidopsis thaliana. Here, we demonstrated that NBR1 (next to BRCA1 gene 1) plays a crucial role as a receptor for selective autophagy during recovery from HS. Immunoblot analysis and confocal microscopy revealed that levels of the NBR1 protein, NBR1-labeled puncta, and NBR1 activity are all higher during the HS recovery phase than before. Co-immunoprecipitation analysis of proteins interacting with NBR1 and comparative proteomic analysis of an nbr1-null mutant and wild-type plants identified 58 proteins as potential novel targets of NBR1. Cellular, biochemical and functional genetic studies confirmed that NBR1 interacts with HSP90.1 (heat shock protein 90.1) and ROF1 (rotamase FKBP 1), a member of the FKBP family, and mediates their degradation by autophagy, which represses the response to HS by attenuating the expression of HSP genes regulated by the HSFA2 transcription factor. Accordingly, loss-of-function mutation of NBR1 resulted in a stronger HS memory phenotype. Together, our results provide new insights into the mechanistic principles by which autophagy regulates plant response to recurrent HS. Abbreviations: AIM: Atg8-interacting motif; ATG: autophagy-related; BiFC: bimolecular fluorescence complementation; ConA: concanamycinA; CoIP: co-immunoprecipitation; DMSO: dimethyl sulfoxide; FKBP: FK506-binding protein; FBPASE: fructose 1,6-bisphosphatase; GFP: green fluorescent protein; HS: heat stress; HSF: heat shock factor; HSFA2: heat shock factor A2; HSP: heat shock protein; HSP90: heat shock protein 90; LC-MS/MS: Liquid chromatography-tandem mass spectrometry; 3-MA: 3-methyladenine; NBR1: next-to-BRCA1; PQC: protein quality control; RFP: red fluorescent protein; ROF1: rotamase FKBP1; TF: transcription factor; TUB: tubulin; UBA: ubiquitin-associated; YFP: yellow fluorescent protein
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