›› 2017, Vol. 60 ›› Issue (11): 1278-1284.doi: 10.16380/j.kcxb.2017.11.005

• 研究论文 • 上一篇    下一篇


王姣玲, 马亚飞, 王霜, 阚云超, 张冰*   

  1. (南阳师范学院, 河南省伏牛山昆虫生物学重点实验室, 昆虫生物反应器河南省工程实验室, 中英南阳洛桑昆虫生物学联合实验室, 河南南阳 473061)
  • 出版日期:2017-11-20 发布日期:2017-11-20

Dynamic distribution of histone H3 Ser10 phosphorylation in Spodoptera frugiperda Sf9 cells during mitosis

WANG Jiao-Ling, MA Ya-Fei, WANG Shuang, KAN Yun-Chao, ZHANG Bing*   

  1. (Henan Key Laboratory of Insect Biology in Funiu Mountain, Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-Res Joint Laboratory of Insect Biology, Nanyang Normal University, Nanyang, Henan 473061, China)
  • Online:2017-11-20 Published:2017-11-20

摘要: 【目的】探讨表观遗传标记组蛋白H3Ser10磷酸化在草地贪夜蛾Spodoptera frugiperda Sf9细胞系细胞有丝分裂中的功能。【方法】序列比对分析组蛋白H3保守性。通过固相法合成特定位点磷酸化修饰的一段组蛋白H3的肽段【RK(pS)TGGKAPRKQLC】进行抗体制备;培养Sf9细胞,通过细胞爬片制备有丝分裂的片子,统计不同时期的细胞数目;以免疫荧光标记检测组蛋白H3Ser10磷酸化抗体在不同时期的定位特点。【结果】序列比对分析发现组蛋白H3第1-60位氨基酸在大多数物种中高度保守。在草地贪夜蛾Sf9细胞中,组蛋白H3Ser10磷酸化发生在早前期细胞的核膜附近,成点状分布;随着细胞周期的进行,早中期达到最高水平,在整条染色体上集中分布。有丝分裂后期开始去磷酸化,并于末期完成去磷酸化。【结论】组蛋白H3Ser10磷酸化与Sf9细胞有丝分裂中染色质的凝缩相关。

关键词: Sf9细胞系; 组蛋白H3; 磷酸化, 有丝分裂, 免疫荧光标记, 染色质凝缩

Abstract: 【Aim】 To explore the function of histone H3 Ser10 phosphorylation, an epigenetic marker, in cells of Spodoptera frugiperda cell line Sf9 during mitosis. 【Methods】 The sequence conservation of histone H3 was analyzed by sequence alignment. Antibody was prepared by synthesis of histone H3 pepetide RK(pS)TGGKAPRKQLC, in which the tenth serine is specifically phosphorylated by solid phase method. Sf9 cells were cultured, and the mitotic cells were prepared by cell climbing. The numbers of cells in different mitotic stages were counted, and then the localization characteristics of histone H3 Ser10 phosphorylation antibody in these stages were detected by cytology immunofluorescence. 【Results】 Sequence alignment analysis revealed that histone H3 1st-60th amino acids were highly conserved in most species. In Sf9 cells, histone H3 Ser10 phosphorylation began to appear near the nuclear membrane with a punctate distribution pattern in early prophase, and spread throughout the whole chromosome in early metaphase during the cell cycle. During anaphase, the fluorescent signals of Ser10-phosphorylated H3 disappeared from chromosomes, and the dephosphorylation completed in telophase. 【Conclusion】 Histone H3 Ser10 phosphorylation is associated with chromosome condensation in Sf9 cells during mitosis.

Key words: Sf9 cell line, histone H3, phosphorylation, mitosis, immunofluorescence labeling, chromosome condensation