The Potential Roles Of The Apoptosis Related Protein PDRG1 In Diapause Embryo Restarting Of Artemia Sinica

Artemia sinica belongs to the arthropods,and is widely distributed in the high salt water body.It has strong resistance to stress and has two kinds of reproductive methods.Eggs produced by the fertilized eggs in the form of dormant eggs excreted in vitro,very low levels of metabolism,resulting in the phenomenon of apoptosis,but dormant eggs for high and low temperature,high salt and other extreme environmental impact,because of its characteristics,has become an experimental biology of developmental biology.P53 and DNA damage-regulated gene 1(PDRG1)is involved in cellular physiological activities,such as apoptosis,DNA damage repair,cell cycle regulation,and promoting programmed cell death.However,the role of pdrg1 in diapause and diapause termination in A.sinica remains unknown.Here,the full-length A.sinica pdrg1 cDNA(As-pdrg1)was obtained and found to contain 1119 nucleotides,including a 228 bp open reading frame(ORF),a 233 bp 5?-untranslated region(UTR),and a 658-bp 3?-UTR,which encodes a 75 amino acid protein.he PDRG1 protein was found to be non-transmembrane protein by TMHMM software.Subcellular localization showed that PDRG1 was a molecule that was mostly located in the nucleus and mitochondria.Informational analysis showed that PDRG1 protein doesn't have a signal peptide sequence,non-secretory protein,belonging to the hydrophilic protein.Phosphorylation sites predicted that there may be 12 phosphorylation sites(score> 0.5)in the Artemia PDRG1 and 8 serine sites.The results of secondary structure prediction showed that PDRG1 was mainly ?-helix(48%),but also had extension chain(18.67%),?-fold(10.67%)and random curl(22.67%).In situ hybridization showed no tissue specific expression of As-pdrg1.Quantitative real-time PCR and western blotting analyses of As-pdrg1 gene and protein expression showed high levels at 15–20 h of embryo development and a subsequent downward trend.Low temperatures upregulated As-pdrg1 expression.RNA interference for the pdrg1 gene in Artemia embryos caused significant developmental hysteresis.Thus,PDRG1 plays an important role in diapause termination and cell cycle regulation in early embryonic development of A.sinica.