Identification of HSF1 Target Genes Involved in Thermal Stress in the Pacific Oyster Crassostrea gigas by ChIP-seq
Identification of HSF1 Target Genes Involved in Thermal Stress in the Pacific Oyster Crassostrea gigas by ChIP-seq
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DOI：
10.1007/s10126-019-09942-6
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发表时间：
2020-04-01
影响因子：
3
通讯作者：
Zhang, Guofan
中科院分区：
文献类型：
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作者：
Liu, Youli;Zhu, Qihui;Zhang, Guofan
太平洋牡蛎Crassostrea Gigas是生活在潮间带的重要商业物种，面临着巨大的温度波动。因此，确定与耐热性相关的候选基因和关键调控关系对牡蛎分子育种具有重要意义。热休克转录因子1(HSF1)在热应激抗性中起重要作用。然而，热休克蛋白(HSP)热休克蛋白(HSP)70和HSF1之间的调控关系尚不清楚。在本研究中，我们通过染色质免疫沉淀和测序(CHIP-SEQ)分析了HSF1在热休克反应中的调控基因，通过qRT-PCR确定了目的基因的表达模式，并验证了一个HSP70和HSF1之间的调控关系。我们发现了与HSF1结合位点相对应的916个峰，并将这些峰注释为最近的基因。在基因本体论分析中，HSF1靶基因与信号转导、能量产生和对生物刺激的反应有关。4个HSP70基因、2个HSP40基因和1个小HSP基因与HSF1结合。在热休克条件下，一个结合在启动子区域的HSP70被证实受HSF1的调控。这些结果为未来旨在确定耐热性的机制的研究提供了基础，并为太平洋牡蛎的基因调控提供了见解。
The Pacific oyster Crassostrea gigas, a commercially important species inhabiting the intertidal zone, facing enormous temperature fluctuations. Therefore, it is important to identify candidate genes and key regulatory relationships associated with thermal tolerance, which can aid the molecular breeding of oysters. Heat shock transcription factor 1 (HSF1) plays an important role in the thermal stress resistance. However, the regulatory relationship between the expansion of heat shock protein (HSP) HSP 70 and HSF1 is not yet clear in C. gigas. In this study, we analyzed genes regulated by HSF1 in response to heat shock by chromatin immunoprecipitation followed by sequencing (ChIP-seq), determined the expression patterns of target genes by qRT-PCR, and validated the regulatory relationship between one HSP70 and HSF1. We found 916 peaks corresponding to HSF1 binding sites, and these peaks were annotated to the nearest genes. In Gene Ontology analysis, HSF1 target genes were related to signal transduction, energy production, and response to biotic stimulus. Four HSP70 genes, two HSP40 genes, and one small HSP gene exhibited binding to HSF1. One HSP70 with a binding site in the promoter region was validated to be regulated by HSF1 under heat shock. These results provide a basis for future studies aimed at determining the mechanisms underlying thermal tolerance and provide insights into gene regulation in the Pacific oyster.