Chilling stress reduced protein translation by the ubiquitination of ribosomal proteins in Volvariella volvacea
Chilling stress reduced protein translation by the ubiquitination of ribosomal proteins in Volvariella volvacea
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DOI：
10.1016/j.jprot.2020.103668
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发表时间：
2020-03-20
影响因子：
3.3
通讯作者：
Bao, Dapeng
中科院分区：
文献类型：
--
作者：
Gong, Ming;Li, Zhengpeng;Bao, Dapeng
草菇是一种重要的食用菌，低温自溶是其代谢异常的一个典型部分，但其机制尚不清楚。泛素组分析表明，冷胁迫（CS）影响蛋白质的翻译和降解的泛素化。比较蛋白质组学分析表明，CS下调草菇V23中的蛋白质表达，而不是VH 3（提高抗冷胁迫的菌株）。整合泛素组，蛋白质组学和转录组学分析表明，CS减少蛋白质翻译的核糖体蛋白的泛素化。20 S蛋白酶体的活性测定表明，CS降低了泛素-蛋白酶体系统的降解效率。蛋白质印迹分析表明，草菇中的泛素结合酶E2（UBE 2）在冷胁迫处理后表达上调。UBEV 2的GST下拉实验提供了证据，CS通过核糖体蛋白的泛素化影响蛋白质翻译。Co-IP实验证实UBEV 2与泛素化的SSB 2结合，SSB 2是一种核糖体相关的分子伴侣。抗冻实验表明，UBE 2抑制剂能提高草菇的抗寒能力。我们的观察结果表明，CS引发的泛素介导的自溶与蛋白质翻译的减少，并强调了UBEV 2在促进低温自溶在V. volvacea.Significance：草菇，食用草菇，是一种高营养的食物来源，广泛种植在热带和亚热带地区的商业规模。与草菇的低温自溶保存相关的挑战限制了其市场化。低温自溶问题既是一个有趣的科学问题，也是一个实际的经济问题。综合泛素组学、蛋白质组学和转录组学分析，以及GST下拉和Co-IP实验表明，低温胁迫通过草菇核糖体蛋白的泛素化降低了蛋白质翻译。这项研究有助于我们理解草菇中泛素化介导的自溶与蛋白质翻译减少相关。我们的数据突出了UBEV 2在促进草菇低温自溶中的机制作用。通过抑制UBEV 2的表达，提高草菇的商品性，为草菇的保存提供了新的思路。
In Volvariella volvacea, an important edible mushroom species, cryogenic autolysis is a typical part of abnormal metabolism; however, the underlying mechanisms remain unclear. Ubiquitylome analysis revealed that chilling stress (CS) affected protein translation and degradation by ubiquitination. Comparative proteomics analysis showed that CS downregulated protein expression in V. volvacea V23 instead of VH3 (improved chilling stress resistance strain). The integrative ubiquitylome, proteomics, and transcriptome analyses indicated that CS reduced protein translation by the ubiquitination of ribosomal proteins. An activity assay of the 20S proteasome showed that CS decreased the degradation efficiency of the ubiquitin-proteasome system. UBEV2, one type of ubiquitin-conjugating enzyme E2 (UBE2) in V. volvacea, was upregulated after cold stress treatment using western blot analysis. GST pull-down experiments of UBEV2 provided evidence that CS affected protein translation by the ubiquitination of ribosomal proteins. Co-IP experiments confirmed that UBEV2 bound to the ubiquitinated SSB2, a ribosome-associated molecular chaperone. An anti-freezing experiment demonstrated that the UBE2 inhibitor could improve the cold stress resistance of V. volvacea. Our observations revealed that CS triggered ubiquitination-mediated autolysis associated with a decrease in protein translation and highlighted the mechanistic role of UBEV2 in facilitating cryogenic autolysis in V. volvacea.Significance: Volvariella volvacea, the edible straw mushroom, is a highly nutritious food source widely cultivated on a commercial scale in tropical and subtropical regions. The challenges associated with the cryogenic autolysis preservation of V. volvacea have limited its marketability. This issue of cryogenic autolysis is both an interesting scientific problem to solve and a practical economic matter. Integrative ubiquitylome, proteomics, and transcriptome analyses, together with GST pulldown and Co-IP experiments, indicated that chilling stress reduced protein translation by the ubiquitination of ribosomal proteins in V. volvacea. This study significantly contributes to our understanding of ubiquitination-mediated autolysis associated with a decrease in protein translation in V. volvacea. Our data highlight the mechanistic role of UBEV2 in facilitating the cryogenic autolysis of V. volvacea. We provided a new idea for the preservation of V. volvacea by inhibiting UBEV2 to increase its marketability.