泥鳅DNA甲基转移酶基因家族结构及时空表达
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作者单位:

1.华中农业大学水产学院,武汉 430070;2.广西壮族自治区水产引育种中心,南宁 530001;3.西华师范大学生命科学学院/西南野生动植物资源保护教育部重点实验室,南充 637009;4.成都师范学院化学与生命科学学院,成都 611130

作者简介:

向玉梅,E-mail: 1353859613@qq.com

通讯作者:

曾燏,E-mail: zengyu_cwnu@163.com
周小云,E-mail: zhouxy@mail.hzau.edu.cn

中图分类号:

S917.4;Q959.46+8

基金项目:

西南野生动植物资源保护教育部重点实验室开放基金(XNYB17-1)


Structure and spatiotemporal expression analysis of DNA methyltransferase gene family of loach Misgurnus anguillicaudatus
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Affiliation:

1.College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China;2.Guangxi Introduction and Breeding Center of Aquaculture, Nanning 530001, China;3.Key Laboratory of Southwest China Wildlife Resources Conservation, Ministry of Education/College of Life Science, China West Normal University, Nanchong 637009, China;4.College of Chemistry and Life Science, Chengdu Normal University, Chengdu 611130, China

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    摘要:

    为深入解析DNA甲基化对泥鳅等硬骨鱼类基因表达调控的作用机制,基于转录组测序数据,用hiTAIL-PCR技术克隆获得泥鳅DNA甲基转移酶基因家族dnmts的启动子序列,并用qRT-PCR和原位杂交技术分析dnmts的时空表达特征。结果显示,维持甲基化酶dnmt1与从头甲基化酶dnmt3s的亚细胞定位、N端调控结构域存在较大的差异,dnmt1的调控域包含DMAP、PBD、CXXC和BAH保守结构域,而dnmt3s包含PWWP、ADD或CH结构域,提示dnmt1dnmt3s间存在较大的功能分化。在4个dnmts的启动子序列上,均预测到典型元件TATA-box,以及CREB、Egr-1、Pit-1等与胚胎发育、生长、繁殖等相关的转录因子结合位点。4个dnmts的表达量均与发育阶段相关、呈动态变化,在卵裂期到囊胚期的快速卵裂阶段,dnmt1高表达,而在胚胎后期的器官分化阶段,dnmt3aadnmt3ab高表达。在成鱼阶段,dnmt1在性腺中高表达,dnmt3aadnmt3ab在脑中高表达,而dnmt3b在性腺、脑和肌肉中均有较高的表达水平。Dnmts的表达呈明显的性别二态性,dnmt1dnmt3b在卵巢中高表达,而dnmt3aadnmt3ab则在精巢中高表达。原位杂交结果显示,dnmt3aa在卵巢的生殖细胞和体细胞中均有表达,而dnmt3ab仅在卵巢的体细胞中表达。以上结果表明,泥鳅dnmts各成员之间存在功能分化,并在特定的发育阶段和组织中发挥重要功能。

    Abstract:

    To analyze the mechanism of DNA methylation in regulating gene expression in teleost such as the loach Misgurnus anguillicaudatus, the promoter sequences of DNA methyltransferase gene family dnmts) of M. anguillicaudatus were cloned using the hiTAIL-PCR method based on transcriptome sequencing data, and the spatiotemporal expression patterns of dnmts were then analyzed using qRT-PCR and in situ hybridization techniques. The results showed significant differences in the subcellular localization and the N-terminal regulatory domain. The regulatory domain of dnmt1 includes the conserved domains DMAP, PBD, CXXC, and BAH, while dnmt3s contain the domains PWWP, ADD, or CH, indicating a notable functional differentiation between dnmt1 and dnmt3s. Such functional differentiation was further confirmed by the phylogenetic tree constructed based on amino acid sequences. In the promoter sequences of the 4 dnmts, besides the typical functional element TATA-box, several transcription factor binding sites related to embryonic development, growth, and reproduction were also predicted, such as CREB,Egr-1 and Pit-1. The expression levels of the 4 dnmts were all associated with the developmental stages and exhibited dynamic changes. During the rapid cleavage stage, from the cleavage stage to the blastocyst stage, dnmt1 was highly expressed, while during the organ differentiation stage in later embryonic development, dnmt3aa and dnmt3ab were highly expressed. In adult fish, dnmt1 was highly expressed in the gonads; dnmt3aa and dnmt3ab were highly expressed in the brain, while dnmt3b showed high expression in the gonads and muscles. Additionally, the expression of dnmts exhibited obvious sex dimorphism: the expression levels of dnmt1 and dnmt3b were significantly higher in the ovaries than in the testis, while dnmt3aa and dnmt3ab showed the opposite pattern. The results of in situ hybridization showed that dnmt3aa was expressed in both germ cells and somatic cells of the ovary, whereas dnmt3ab was expressed only in the somatic cells of the ovary. These findings indicate functional differentiation among the members of the dnmts family in M. anguillicaudatus, suggesting that they play important roles in specific developmental stages and tissues.

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向玉梅,苏君晓,曾燏,周小云.泥鳅DNA甲基转移酶基因家族结构及时空表达[J].华中农业大学学报,2026,45(1):255-265

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  • 收稿日期:2024-03-04
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  • 在线发布日期: 2026-02-09
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