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基于液相色谱-质谱联用技术的模式触发免疫中核蛋白质组和蛋白质导入的草图图谱

LC-MS Based Draft Map of the Nuclear Proteome and Protein Import in Pattern Triggered Immunity.

作者信息

Ayash Mohamed, Abukhalaf Mohammad, Thieme Domenika, Proksch Carsten, Heilmann Mareike, Schattat Martin Hartmut, Hoehenwarter Wolfgang

机构信息

Department Biochemistry of Plant Interactions, Leibniz Institute of Plant Biochemistry, Halle, Germany.

Institute for Biochemistry and Biotechnology, Martin-Luther University Halle-Wittenberg, Halle, Germany.

出版信息

Front Plant Sci. 2021 Nov 8;12:744103. doi: 10.3389/fpls.2021.744103. eCollection 2021.

Abstract

Despite its central role as the ark of genetic information and gene expression the plant nucleus is surprisingly understudied. We isolated nuclei from the dark grown cell culture left untreated and treated with flg22 and nlp20, two elicitors of pattern triggered immunity (PTI) in plants, respectively. An liquid chromatography mass spectrometry (LC-MS) based discovery proteomics approach was used to measure the nuclear proteome fractions. An enrichment score based on the relative abundance of cytoplasmic, mitochondrial and Golgi markers in the nuclear protein fraction allowed us to curate the nuclear proteome producing high quality catalogs of around 3,000 nuclear proteins under untreated and both PTI conditions. The measurements also covered low abundant proteins including more than 100 transcription factors and transcriptional co-activators. We disclose a list of several hundred potentially dual targeted proteins including proteins not yet found before for further study. Protein import into the nucleus in plant immunity is known. Here we sought to gain a broader impression of this phenomenon employing our proteomics data and found 157 and 73 proteins to possibly be imported into the nucleus upon stimulus with flg22 and nlp20, respectively. Furthermore, the abundance of 93 proteins changed significantly in the nucleus following elicitation of immunity. These results suggest promiscuous ribosome assembly and a role of prohibitins and cytochrome C in the nucleus in PTI.

摘要

尽管植物细胞核作为遗传信息和基因表达的核心载体发挥着关键作用,但令人惊讶的是,对其研究却相对不足。我们从未经处理以及分别用flg22和nlp20处理过的黑暗培养的细胞培养物中分离细胞核,flg22和nlp20是植物中模式触发免疫(PTI)的两种激发子。采用基于液相色谱质谱(LC-MS)的发现蛋白质组学方法来测量细胞核蛋白质组组分。基于细胞核蛋白组分中细胞质、线粒体和高尔基体标记物的相对丰度的富集分数,使我们能够精心挑选细胞核蛋白质组,生成在未处理以及两种PTI条件下约3000种细胞核蛋白的高质量目录。这些测量还涵盖了低丰度蛋白质,包括100多种转录因子和转录共激活因子。我们公布了数百种潜在的双靶向蛋白列表,其中包括一些此前尚未发现的蛋白以供进一步研究。植物免疫中蛋白质向细胞核的导入是已知的。在此,我们试图利用我们的蛋白质组学数据对这一现象有更广泛的认识,发现分别用flg22和nlp20刺激后,可能有157种和73种蛋白质会导入细胞核。此外,免疫激发后细胞核中93种蛋白质的丰度发生了显著变化。这些结果表明在PTI中,核糖体组装杂乱,并且禁止素和细胞色素C在细胞核中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b0a/8630587/ce506fafcbb1/fpls-12-744103-g001.jpg

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