Structure and spatiotemporal expression analysis of DNA methyltransferase gene family of loach Misgurnus anguillicaudatus
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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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S917.4;Q959.46+8

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    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. Structure and spatiotemporal expression analysis of DNA methyltransferase gene family of loach Misgurnus anguillicaudatus[J]. Jorunal of Huazhong Agricultural University,2026,45(1):255-265.

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History
  • Received:March 04,2024
  • Revised:
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  • Online: February 09,2026
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