The Fuction Of INTS11 In Diapause Embryo Restart Of Artemia Sinica

Artemia sinica can produce protective diapause eggs in adverse environments.The diapause can be broken by appropriate conditions.The molecular regulation mechanism of Artemia sinica is still not clear.Through related research,it is found that the integrator complex subunit 11 plays an indispensable role in the process of embryonic resuscitation,especially in the cell cycle regulation.However,the mechanism of integration factor complex 11 in the process of diapause of Artemia sinica diapause embryos is still unknown.Integrator complex subunit 11 is an endonuclease that interacts with the C-terminus of a repeating amino acid sequence of the RNA polymerase II(RANPII)large subunit,which in turn mediates the 3 ' end of the small nuclear RNA and enhancer RNA(eRNA)processing,it is also possible to participate in the cleavage of the pre-mRNA encoding the protein required for G1 progression and entry into the S phase,so as to form mature mRNA and promote transcription of the target gene in the cell cycle.Therefore,the integrator complex subunit 11 may play an important role in the re-initiation of Artemia in China.This study explores the theoretical analysis and experimental exploration of the role of the integrator complex subunit 11 in Artemia sinica.In this study,a 1964 bp full-length cDNA of integrator complex subunit from Artemia sinica was cloned for the first time,it encodes 597 amino acids,a hydrophobic protein with a molecular weight of about 67 KDa,and the isoelectric point is 6.79;Subcellular localization prediction showed that the integrator complex subunit 11 is most likely to be highly expressed in the nucleus and also in the cytoplasm;The protein detected by SignalP4.1 does not contain a signal peptide,and the integrator complex subunit 11 belongs to a non-secreted protein;it is predicted by SMART that the protein contains two domains of metallo-hydrolase-like-MBL-fold and Beta-casp;The gene and protein expression levels of the integrator complex subunit 11 in Artemia sinica were detected by qPCR and Western blotting.The results showed that the gene and protein levels were consistent,and the expression increased during the 0-15 h development period,and the expression gradually increased after 15 h.Lower,indicating that theintegrator complex subunit 11 may play an important role in the embryo restart process,However,in temperature and salinity stress,the INTS11 has the lowest expression at low temperature and high salt,and when the environment is suitable,its expression level increases,indicating that the integrator complexsubunit 11 is also affected by temperature during the regulation of embryo development regulation of salinity.At the same time,Western blotting was used to detect the expression levels of INTS11-related proteins INTS9 and RANTII.The results showed that they were consistent with the expression trend of INTS11.It is speculated that INTS11 and INTS9 may form heterodimers during embryonic development and then with the C-terminus of RANTII.In combination with the splicing of mRNA,it promotes the transcription of cell cycle target genes.This paper further analyzes the molecular mechanism of integration factor complex 11 in the reactivation of Artemia sinica diapause embryos,and provides a scientific basis for the study of the molecular mechanism of crustacean and other invertebrate diapause embryos.