Characterization of the core region of grape VvHOS1 promoter activity and its upstream regulatory proteins
Characterization of the core region of grape VvHOS1 promoter activity and its upstream regulatory proteins
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葡萄VvHOS1启动子活性核心区及其上游调控蛋白的表征
DOI：
10.1016/j.envexpbot.2022.105199
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发表时间：
2023-01-02
影响因子：
5.7
通讯作者：
Xu，Weirong
中科院分区：
文献类型：
--
作者：
Wu，Nan;Li，Junduo;Xu，Weirong
葡萄(VitisviniferaL.)的热带起源使其容易受到低温的影响，这对葡萄生产构成了严重威胁。我们对多年生葡萄耐冷性的分子基础的了解是有限的。渗透反应基因1(HOS1)的E3泛素连接酶高表达已被证明对冷反应具有负性调节作用，但对HOS1在葡萄中的了解还不是很清楚。在这里，我们初步评估了HOS1在耐冷山葡萄(VaHOS1)和敏感葡萄(VaHOS1)中的表达。VvHOS1)品种，发现它们对冷胁迫的反应存在显著差异。两个葡萄HOS1基因具有很高的序列相似性(在氨基酸水平上为98.77%)，但它们的长度(313个碱基插入/缺失多态)和启动子序列显著不同(91.96%)。这一313个碱基的区域显著影响HOS1在V和V之间的差异表达。Vinifera和V。阿穆尔山。在转基因拟南芥启动子-GUS分析中，313个碱基的缺失取消了VvHOS1启动子的活性。此外，313bp的片段可能是影响VvHOS1to启动子活性的正核心区，参与了不同发育阶段和冷胁迫的过程。此外，DNA下拉和质谱分析表明，潜在的调控蛋白可能结合在313-bp的核心启动子区域。我们发现，两个连接物组蛋白H1基因(命名为VvHH1：LOC100853898和LOC100259836)和一个姐妹染色单体凝聚蛋白Pds5同源物(命名为VvPDS5B：LOC100853350)与核心启动子区域相关，降低或激活GUS的表达。这项工作阐明了HOS1启动子在基因组中的表达特征，并进一步确定了通过激活或抑制候选调节蛋白来激活或抑制启动子活性所需的核心区域。
The tropical origins of the grape (VitisviniferaL.) make it vulnerable to low temperatures, which constitute a serious threat to grape production. Our knowledge of the molecular basis of cold tolerance in perennial grape species is limited. The E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE 1 (HOS1) has been demonstrated to negatively regulate cold responsiveness, but the understanding of theHOS1in grape remains fragmentary. Here, we initially evaluated the expression ofHOS1in cold-tolerantVitis amurensis(VaHOS1) and susceptibleV. vinifera(VvHOS1) cultivars and discovered their responses to cold stress differed significantly. The two grapeHOS1genes exhibit a high degree of sequence similarity (98.77% at the amino acid level) but their lengths (313-bp insertion/deletion polymorphism) and promoter sequences are substantially different (91.96%). This 313-bp region significantly affects the differential expression ofHOS1betweenV. viniferaandV. amurensis. The 313-bp deletion abolishedVvHOS1promoter activity in promoter-GUS assay of transgenicArabidopsis. Moreover, 313-bp fragment might function as a positive core region to affect promoter activity ofVvHOS1to involved in different developmental stages and cold stress. In addition, DNA pull-down and mass spectrometry (MS) assay revealed the potential regulatory proteins may bind to the 313-bp core promoter region. We show that the two linker histone H1 genes (designated asVvHH1: LOC100853898 and LOC100259836), and one sister chromatid cohesion protein PDS5 homolog B (designated asVvPDS5B: LOC100853350), which associates with the core promoter region, decreases or activatesGUSexpression. This work elucidates the expression features of theHOS1promoter from genusVitisand further identifies a core region required for the promoter activity viatrans-activation or suppression of candidate regulatory proteins.