College of Life Sciences, Liaoning Normal University, Dalian, PR China.
Department of Biology, Dalian Medical University, Dalian, PR China.
PLoS One. 2014 Jan 3;9(1):e85343. doi: 10.1371/journal.pone.0085343. eCollection 2014.
During embryonic development of Artemia sinica, environmental stresses induce the embryo diapause phenomenon, required to resist apoptosis and regulate cell cycle activity. The small ubiquitin-related modifier-1 (SUMO), a reversible post-translational protein modifier, plays an important role in embryo development. SUMO regulates multiple cellular processes, including development and other biological processes. The molecular mechanism of diapause, diapause termination and the role of As-sumo-1 in this processes and in early embryo development of Artemia sinica still remains unknown. In this study, the complete cDNA sequences of the sumo-1 homolog, sumo ligase homolog, caspase-1 homolog and cyclin B homolog from Artemia sinica were cloned. The mRNA expression patterns of As-sumo-1, sumo ligase, caspase-1, cyclin B and the location of As-sumo-1 were investigated. SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E proteins were analyzed during different developmental stages of the embryo of A. sinica. Small interfering RNA (siRNA) was used to verify the function of sumo-1 in A. sinica. The full-length cDNA of As-sumo-1 was 476 bp, encoding a 92 amino acid protein. The As-caspases-1 cDNA was 966 bp, encoding a 245 amino-acid protein. The As-sumo ligase cDNA was 1556 bp encoding, a 343 amino acid protein, and the cyclin B cDNA was 739 bp, encoding a 133 amino acid protein. The expressions of As-sumo-1, As-caspase-1 and As-cyclin B were highest at the 10 h stage of embryonic development, and As-sumo ligase showed its highest expression at 0 h. The expression of As-SUMO-1 showed no tissue or organ specificity. Western blotting showed high expression of As-SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E at the 10 h stage. The siRNA caused abnormal development of the embryo, with increased malformation and mortality. As-SUMO-1 is a crucial regulation and modification protein resumption of embryonic diapause and early embryo development of A. sinica.
在卤虫胚胎发育过程中，环境压力会诱导胚胎休眠现象，这是抵抗细胞凋亡和调节细胞周期活动所必需的。小泛素相关修饰物-1（SUMO）是一种可逆的翻译后蛋白修饰物，在胚胎发育中发挥着重要作用。SUMO 调节多种细胞过程，包括发育和其他生物过程。休眠、休眠终止的分子机制以及 As-sumo-1 在这些过程以及卤虫早期胚胎发育中的作用尚不清楚。在这项研究中，克隆了卤虫 sumo-1 同源物、sumo 连接酶同源物、caspase-1 同源物和 cyclin B 同源物的全长 cDNA 序列。研究了 As-sumo-1、sumo 连接酶、caspase-1、cyclin B 的 mRNA 表达模式及其在卤虫胚胎不同发育阶段的定位。分析了 SUMO-1、p53、Mdm2、Caspase-1、Cyclin B 和 Cyclin E 蛋白在卤虫胚胎不同发育阶段的表达。利用小干扰 RNA（siRNA）验证了 sumo-1 在卤虫中的功能。As-sumo-1 的全长 cDNA 为 476bp，编码 92 个氨基酸的蛋白质。As-caspases-1 cDNA 为 966bp，编码 245 个氨基酸的蛋白质。As-sumo 连接酶 cDNA 为 1556bp，编码 343 个氨基酸的蛋白质，cyclin B cDNA 为 739bp，编码 133 个氨基酸的蛋白质。As-sumo-1、As-caspase-1 和 As-cyclin B 的表达在胚胎发育的 10h 阶段最高，而 As-sumo 连接酶在 0h 时表达最高。As-SUMO-1 的表达没有组织或器官特异性。Western blot 显示，在 10h 阶段，As-SUMO-1、p53、Mdm2、Caspase-1、Cyclin B 和 Cyclin E 的表达水平较高。siRNA 导致胚胎发育异常，畸形率和死亡率增加。As-SUMO-1 是调节卤虫胚胎休眠和早期胚胎发育恢复的关键调控和修饰蛋白。
