The Potential Role Of CCAR-1 During The Diapauses Embryonic Restarting Process Of Artemia Sinica

Artemia sinica is a small crustacean that endure high salinity and wide temperature range,widely distributed habitants in both inland and coastal saltworks and it is with strong resistance to environmental stress.With the survival environment degradation,a diapause stage will occur during the process of embryo development.When the environmental conditions was suitable,diapause would be terminated,and embryonic development would restar.Cell division cycle and apoptosis regulator protein 1(CCAR-1)plays an important role in the process of cell cycle.It can bind other cell cycle regulatory proteins which promote cell apoptosis.CCAR-1 and ?-catenin were activated which assist it transcriptional activation to the target genes in Wnt parthway.CCAR-1 combin with APC-2,a subunit of APC/C,repress cells to enter in G2/M,phase under condition of the activation of CFM-4.It can participate in caspase ubiquitin proteasome pathway,resulting in a deletion of cyclinB1.CCAR-1compound with P53 as an activation agent,promotes the process of transcription regulation and as an apoptosis signal to regulate of cell cycle.CCAR-1 compose with p160 an activator compounds and it can produce hormone on the appropriate target gene in the nuclear.Therefore,it plays an important role in the process of cell cycle regulation.But the function of CCAR-1 in cell cycle of diapause termination from Artemia sinica still remains unknown.In the present study,partial sequnce of cDNA from As-CCAR-1 of A.sinicain was obtained by the method of RACE,Genome Walking,and bioinformatics analysis was performed.The gene expression levels of CCAR-1 sequence at different developmental stages,low temperature,high salinity stress,were analyzed by Real-Time qPCR,insitu hybridization and immunofluorescence.The cDNA sequnce obtained of As-CCAR-1 is 1594 bp and an open reading frame of1584 bp which contains three domain,S1-LIKE,DBC1 and SAP,theoretical pl/Mw:4.99/129382.70,hydrophobic.The result of subcellular localization shows that CCAR-1 distributes in the whole body and cells.Real-Time qPCR results the expression level of the highest in 3d after diapause termination,expression level significantly reduced under low temperature and high salt stress.Above all the experimental result,showed that CCAR-1is associated with cell cycle and cell apoptosis in embryo diapause termination of in A.sinica.The study was to clarify the function of As-CCAR-1 in the process of cell cycle regulation and developmet after diapause relieved.It has important significance for further to know of role of As-CCAR-1 in diapause ternination from A.sinica.