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马氏珠母贝组蛋白H1基因家族的鉴定及功能分析

Identification and Functional Differentiation Analysis of Histone H1 Gene Family in Pinctada fucata martensii

  • 摘要: 组蛋白H1参与高级染色质结构的形成,除结构作用外,组蛋白H1还通过翻译后修饰或特定变体等影响染色质功能和动力学,并具有参与生物免疫反应以及控制细胞增殖等功能。本研究通过构建系统进化树和多序列比对等方法比较分析了马氏珠母贝(Pinctada fucata martensii)、长牡蛎(Crassostrea gigas)和人(Homo sapiens)等9个物种的组蛋白H1基因家族,分析了马氏珠母贝中的组蛋白H1基因在不同发育阶段以及植核后不同时间血细胞转录组中的表达变化,旨在揭示马氏珠母贝中组蛋白H1基因家族的分子进化和功能。结果表明,马氏珠母贝基因组包含23个组蛋白H1基因变体,呈现了该生物的特异性扩张。对马氏珠母贝、长牡蛎和人的组蛋白H1的氨基酸序列进行多序列比对,结果显示,组蛋白H1在物种间的保守性较高。在马氏珠母贝不同发育阶段的转录组中检测到12个组蛋白H1基因转录本,3个组蛋白H1基因在原肠胚期、早期担轮幼虫期、D型幼虫期、担轮幼虫、D型幼虫、投饵前D型幼虫、早期壳顶幼虫期、眼点期、变态后时期和稚贝期表达量升高,1个组蛋白H1基因在卵期和受精卵期显著性高表达,说明组蛋白H1在发育过程中形成了不同的表达机制,产生功能分化,在不同的发育阶段由不同H1基因发挥不同的功能。在血细胞转录组中表达了11个组蛋白H1基因,在植核后3~6 d有9个组蛋白H1基因的表达量有明显增加,在植核后6~12 h,有2个组蛋白H1基因显著性高表达,在植核后12 d其组蛋白H1基因的表达量都基本恢复到植核前水平,进一步证实不同的组蛋白H1基因的功能分化特征。上述结果表明组蛋白H1基因在马氏珠母贝基因组中产生了特异性扩张并形成功能分化,参与调控马氏珠母贝的发育和植核免疫过程。本研究为进一步探究组蛋白H1基因在马氏珠母贝中的功能提供理论基础。

     

    Abstract: Histone H1 participates in the formation of high-level chromatin structure. In addition to structuralfunction, histone H1 also affects chromatin function and dynamics through the presence of post-translational histone modifications or specific histone variants, and has the ability to participate in biological immune responses and control cell proliferation and other functions. In this study, the histone H1 genes from 9 species, including Pinctada fucata martensii, Crassostrea gigas, and Homo sapiens et al., were identified and analyzed through construction of phylogenetic trees and multiple sequences alignment methods. The expression pattern of H1 gene during the development and after transplantation in Pinctada fucata martensii were re-analyzed to obtain its molecular evolution and functional differentiation characteristics in P.f.martensii. The results showed that the genome of Pinctada martensii contained 23 histone H1 gene variants, showing the specific expansion of the organism. Multi-sequence alignment analysis of the amino acid sequences of the histone H1 gene of Pinctada fucata martensii, Crassostrea gigas and Homo sapiens showed that the sequence of histone H1 was highly conserved among species. Twelve transcripts of histone H1 genes were detected in the transcriptomes at different developmental stages of P.f.martensii. Three histone H1 genes were found in gastrula, early trochophore, trochophore, D-larvae, D-shaped larvae before feeding, early umbo larvae, eye-larvae, post-veliger and juveniles. And one histone H1 gene was highly expressed in the egg stage and the fertilized egg stage. Histone H1 gene had formed different expression mechanisms in the course of evolution, resulting in functional differentiation, which was induced at different developmental stages to perform different functions. A total of 11 histone H1 genes were identified in the transcriptome of the hemocytes. After allograft and xenograft transplantation, the expression of 9 histone H1 genes increased significantly at 3 to 6 days, and 2 histone H1 genes were highly expressed at 6~12 h, and was basically recovered to the level before nucleation at 12 days, which further illustrated the functional differentiation characteristics of different histone H1 gene. The above results indicated that histone H1 had a specific expansion and functional differentiation in the P.f.martensii genome, and was involved in the regulation of the development and nuclear immune process of P.f.martensii. This study provides a theoretical basis for further exploring the function of histone H1 gene in P.f.martensii.

     

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