A VASt-domain protein regulates autophagy, membrane tension, and sterol homeostasis in rice blast fungus.
A VASt-domain protein regulates autophagy, membrane tension, and sterol homeostasis in rice blast fungus.
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VASt 结构域蛋白调节稻瘟病菌的自噬、膜张力和甾醇稳态。
DOI：
10.1080/15548627.2020.1848129
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发表时间：
2021
期刊：
影响因子：
13.3
通讯作者：
Liu,Xiao-Hong
中科院分区：
文献类型：
--
作者：
Zhu,Xue-Ming;Li,Lin;Cai,Ying-Ying;Wu,Xi-Yu;Shi,Huan-Bin;Liang,Shuang;Qu,Ying-Min;Naqvi,NaweedI;DelPoeta,Maurizio;Dong,Bo;Lin,Fu-Cheng;Liu,Xiao-Hong
甾醇是一类对基本生物过程和膜动力学至关重要的脂质。这些分子在内质网（ER）中合成，并在ER和质膜（PM）之间双向转运。然而，甾醇的运输机制及其与自噬/自噬的关系仍然知之甚少。在这里，我们确定了VAD 1类似物的StAR相关的脂质转移（VASt）结构域含有蛋白MoVast 1通过免疫共沉淀在M。米。MoVAST 1的缺失导致分生孢子缺陷，附着胞发育受损，致病性降低。MoTor（M.由于MoVast 1缺失导致PM中高水平的甾醇积累，因此MoVast 1的活性受到抑制。定点突变表明902 T位点对MoVast 1的定位和功能至关重要。通过filipin或Flipper-TR染色，自噬通量检测，MoAtg 8脂质化和药物敏感性测定，我们发现MoVast 1作为一种新的自噬抑制因子，通过调节甾醇含量来监测PM中的张力，从而调节MoTor的活性。脂质组学和转录组学分析进一步证实MoVast 1是脂质代谢和自噬途径的重要调节剂。我们的研究结果揭示了一种新的甾醇转移蛋白的特征。缩写：AmB：异甘草素B; ATMT：根癌农杆菌介导的转化; CM：完全培养基; dpi：接种后天数; ER：内质网; Flipper-TR：荧光脂质张力报告基因; GO：基因本体; hpi：接种后小时数; IH：侵入菌丝; KEGG：基因和基因组的京都百科全书; MoTor：Magnaporthe mesium中雷帕霉素的靶标; PalmC：棕榈酰肉碱; PM：质膜; SD-N：不含氨基酸和硫酸铵的合成成分确定培养基; TOR：雷帕霉素靶标; VASt：StAR相关脂质转移的VAD 1类似物; YFP，黄色荧光蛋白。
Sterols are a class of lipids critical for fundamental biological processes and membrane dynamics. These molecules are synthesized in the endoplasmic reticulum (ER) and are transported bi-directionally between the ER and plasma membrane (PM). However, the trafficking mechanism of sterols and their relationship with macroautophagy/autophagy are still poorly understood in the rice blast fungusMagnaporthe oryzae. Here, we identified the VAD1 Analog of StAR-related lipid transfer (VASt) domain-containing protein MoVast1 via co-immunoprecipitation inM. oryzae. Loss ofMoVAST1resulted in conidial defects, impaired appressorium development, and reduced pathogenicity. The MoTor (target of rapamycin inM. oryzae) activity is inhibited because MoVast1 deletion leads to high levels of sterol accumulation in the PM. Site-directed mutagenesis showed that the 902 T site is essential for localization and function of MoVast1. Through filipin or Flipper-TR staining, autophagic flux detection, MoAtg8 lipidation, and drug sensitivity assays, we uncovered that MoVast1 acts as a novel autophagy inhibition factor that monitors tension in the PM by regulating the sterol content, which in turn modulates the activity of MoTor. Lipidomics and transcriptomics analyses further confirmed that MoVast1 is an important regulator of lipid metabolism and the autophagy pathway. Our results revealed and characterized a novel sterol transfer protein important forM. oryzaepathogenicity.Abbreviations:AmB: amphotericin B; ATMT:Agrobacterium tumefaciens-mediated transformation; CM: complete medium; dpi: days post-inoculation; ER: endoplasmic reticulum; Flipper-TR: fluorescent lipid tension reporter; GO: Gene ontology; hpi: hours post-inoculation; IH: invasive hyphae; KEGG: kyoto encyclopedia of genes and genomes; MoTor: target of rapamycin inMagnaporthe oryzae; PalmC: palmitoylcarnitine; PM: plasma membrane; SD-N: synthetic defined medium without amino acids and ammonium sulfate; TOR: target of rapamycin; VASt: VAD1 Analog of StAR-related lipid transfer; YFP, yellow fluorescent protein.