The Integrator complex (INT) contains several subunits that participate in RNAPII transcription and the 3' end process of non-coding RNAs. INTS11 is the catalytic subunit that interacts with the C-terminal domain of RNAPII, recently found to play a role in embryo development in different experimental models. However, the involvement of INTS11 is still ignorant in crustaceans, particularly in post-diapause embryonic development of Artemia sinica. In the present research, the full-length cDNA of As-Ints11 gene (1964 bp) was cloned from A. sinica by the RACE technique. The deduced 597 amino acids sequence contains the most identifiable domains of the INTs and is highly conserved. Immunofluorescence assay showed that the INTS11 was present at diverse developmental status in A. sinica: the As-INTS11 can be found in both cytoplasm and nucleus of the embryos, and the location showed no specificity in tissue or organ of the nauplius. The expression patterns of As-Ints11 were analyzed by qPCR and Western blotting, which show similar trends that peaked at the 15 h stage of embryo development. Moreover, the expressions of interacting proteins As-INTS9 and As-RNAPII were also detected, results display a synergetic effect with the As-INTS11 at both mRNA and protein levels. We also explored the amount of As-INTS11, As-INTS9 and As-RNAPII under different stresses, and the results indicate that the As-INTS11 is a stress-related protein though the mechanism needs further research. Knocking down of the As-INTS11 resulted in a delay of post-diapaused embryonic development in A. sinica.
Keywords: Artemia sinica; As-Ints11; Diapause; Embryo development; INTS11; Integrator complex.
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