Interaction Of Protein Kinase Pkmyt1 With RASAL2 And Its Effect On The Early Development Of Mouse Embryos

ObjectiveConfirming the interaction between protein kinase Pkmyt1 and RASAL2 during the one-cell stage mouse embryos,exploring the effect on the development of mouse embryos and laying the foundation for further exploring the mechanism of Pkmyt1 in early developmental regulation mammalian embryos.Methods1.CO-IP was used for verifying the interaction between protein kinase Pkmyt1 and RASAL2 in mammalian cell.2.GST pull down was used for verifying the interaction between protein kinase Pkmyt1 and RASAL2 in vitro.3.Immunofluorescence was used for observing the situation of protein kinase Pkmyt1 and RASAL2 in one-cell stage mouse embryos.4.Real-time PCR was used for detecting the expression of RASAL2 mRNA in one-cell stage mouse embryos.5.Immunofluorescence was used for observing the situation of RASAL2 in one-cell stage mouse embryos.6.One-cell stage mouse embryos microinjectioned with RASAL2-mRNA and observed the effect of cleavage rate on the one-cell stage mouse embryos.Results1.Protein kinase Pkmyt1 was interacted with RASAL2 in the cells.2.Protein kinase Pkmyt1 was interacted with RASAL2 in vitro.3.Protein kinases Pkmyt1 and RASAL2 co-localized in one-cell stage mouse embryos.4.RASAL2 mRNA was expressed in one-cell stage mouse embryos.The expression increased in S phase and decreased in G2 phase.5.RASAL2 was mainly located in the cytoplasm of the one-cell stage mouse embryos,and the M phase was expressed in the cytoplasm and nucleus.6.Overexpression of RASAL2 inhibits the cleavage rate of one-cell stage mouse embryos.ConclusionsThis study confirmed that protein kinase Pkmyt1 was interacted with RASAL2,and co-located in one-cell stage mouse embryos.RASAL2 mRNA expression in one-cell stage mouse embryos and overexpression of RASAL2 inhibited the cleavage rate of one-cell stage mouse embryos,it suggest that protein kinase Pkmyt1 may interact with RASAL2 in mouse embryos to regulate early development.