Research On The Reguation Of MSI60 And Prismalin-14 By Pf-Rel In Pinctada Fucata

Pinctada fucata is mainly used to produce pearls in aquaculture.Shells and pearls are typical biomineralized products.Many experiments showed that matric proteins played important roles in shell formation.Thus,the research about matrix proteins has always been a research hotspot.However,researches about upstream regμlatory mechanisms are still limited.NF-кB signaling pathway plays an important role in connecting skeletal system and immune system.Pf-Rel belongs to Rel/ NF-кB family,considered to be the key transcription factor,which can come into the nucleus,combine with DNA,and regulate downstream gene expression.Prismalin-14 and MSI60 mainly function in the formation of prismatic layer and pearl layer.This study focused on the regulation mechanism of Prismalin-14 and MSI60,and we explored the activation sites of these two matrix protein promoters.In this study,we found the relative expression of Prismalin-14 and MSI60 genes were all high in mantle tissue.What’s more,we found thess three genes all expressed higher after shell notching and existed rule of time,suggesting that they all functioned in the shell formation.Then,we silenced the Pf-Rel gene using RNAi method and found the relative expression of these three genes all decreased.At the same time,the promoters of Prismalin-14 and MSI60 were cloned into luciferase reporter vectors and co-transfected with the Pf-Rel gene into HEK-293 T cells.We found there existed obvious dose effect and proved that Pf-Rel transcription factor could activate Prismalin-14 and MSI60 promoters.Finally,we explored the activation site of Prismalin-14 promoter was in the region of-1794 to-1599 bp and the activation site of the MSI60 promoter was in the region of-2244 to-1141 bp.In a summary,transcription factor Pf-Rel can activate Prismalin-14 and MSI60 promoters to regulate the expression of these two matrix proteins.