Characterization And Functional Analysis Of Zfp3612in Early Development

RNA-binding proteins are key components in RNA metabolism. They regulate all aspects of RNA biogenesis from RNA maturation, nucleocytoplasmic transport to subcellular localization. Zfp3612is a member of CCCH tandem zinc finger protein family which contains a zinc finger domain. In mice it has been found that Zfp3612is closely linked with hematopoietic function and cancer, while its roles in early development are not clear. In this study, the characterization and function exploration of Zfp3612in Danio rerio was reported.The full-length of zebrafish Zfp3612cDNA coding sequence is1122bp and encodes a predicted373amino acid protein which is63amino acids more compared to the prediction on NCBI. The molecular weight of Zfp3612protein is41.6kDa and the pI is7.978. Zfp3612has a Tis11B_N in N-terminal followed by two tandem zinc finger domains and Ser/Pro rich region in C-terminal. The tandem zinc finger domain is more conserved compare to Tisl1B_N domain and the C-terminal.Although zebrafish Zfp3612is much shorter than vertebrate Zfp3612, syntenic analysis demonstrated that Zfp3612is an ortholog not a paralog of vertebrate Zfp3612. Phylogenetic analysis revealed that all Zfp36gene family members were originated from an ancient gene, and this occurred before the divergence of fish and vertebrate.Zfp3612is widely expressed in many tissues:eyes, brain, intestines, liver, gill, muscle, heart, and skin, and mainly expressed in brain, liver, heart. Zfp3612is also expressed in all the stages of development. In situ hybridization result suggest that Zfp3612has maternal expression, and it has ubiquitous expression throughout all the early stages, while it mainly exptressed on the head later than1dpf. This may suggest that Zfp3612perform many functions in different cells and at different stages.Gene knockdown/knockout is a good method to study gene function. Morpholino can knockdown gene expression by blocking translation. Knockdown of Zfp3612led to dorsalized embryos with enlarged head and truncated tail. In addition, luciferase experiment revealed that Zfp3612was able to regulate Wnt signal pathway, Taken together, these preliminary results suggest that Zfp3612may play a role in axis formation, possibly by targeting RNAs involved in canonical Wnt singal pathway.In this study, the function of Zfp3612and the mode of action were explored, it was found that Zfp3612involved in axis formation, possibly through Wnt signal pathway. These results may provide novel insight into the mechanisms underlying axis formation.