Abstract
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.
Highlights
Artemia sinica, a small aquatic crustacean, lives in the hyperosmotic environment of salt pools and salt lakes in China [1]
Our aim was to further our understanding of the function of As-sumo-1 and the other proteins in regulation and modification of the cell cycle and apoptosis during postdiapause and in early embryo developmental stages of A. sinica
SignalP3.0 analysis showed that As-small ubiquitin-related modifier (SUMO)-1 has no signal peptide
Summary
A small aquatic crustacean, lives in the hyperosmotic environment of salt pools and salt lakes in China [1]. A. sinica, as an experimental material, is widely used in many fields including physiology, developmental biology, evolution and ecology [2]. Post-translational modifications are involved many cellular processes, such as signal transduction, protein localization and the cell cycle [3]. Phosphorylation, methylation and other modifications by small molecules act as post-translational modifiers. One of the best known modifiers is ubiquitin, which mediates degradation of target proteins by the 26S proteasome [4]. A number of small proteins, classified as ubiquitin-like modifiers (Ubls), have been identified to be covalently attached to target proteins in a similar manner to ubiquitylation. The small ubiquitin-related modifier (SUMO) was defined as a posttranslational modifier following the identification of the first SUMO gene (SMT3) and the first substrate (RanGAP1, Ran GTPase-activating protein 1) [5,6]
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