Identification and functional characterization of SIMPL in Crassostrea gigas reveals its ancient origin and involvement in the regulation of Rel/NF-κB transcription activity

SIMPL (Signaling Molecule that associates with the mouse Pelle-Like Kinase) has been recently identified as a co-regulator of NF-κB dependent transcription. Here, we report the discovery and functional analysis of the SIMPL in a mollusk, Crassostrea gigas, which terms as CgSIMPL. CgSIMPL is comprised of 252 amino acids and shares significant homology with vertebrate homologs. Over-expression of CgSIMPL does not activate the NF-κB reporter in the HEK293 cell line, but can enhance Rel-dependent NF-κB transactivation. The dominant-negative effect of CgSIMPL was observed after the deletion of NLS, strongly suggesting that NLS is required for the enhancement of Rel-dependent NF-κB transactivation. Furthermore, CgSIMPL mRNA is constitutively expressed in various tissues and is inducible at late stages of infection, supporting its regulatory function in innate immunity. Altogether, our studies reveal that SIMPL is reserved and co-evolved with NF-κB in chordate and mollusk, supporting its ancient origin and involvement in regulation of NF-κB signaling pathway.