Transcription factor Pf-Rel regulates expression of matrix protein genes Prismalin-14 and MSI60 in the pearl oyster Pinctada fucata.

Abstract

Molluscan shell is a biomineral that consists of a highly organized calcium carbonate composite. Organisms mainly use matrix proteins to elaborately control the biomineralization process, but knowledge of their regulatory mechanisms is limited. The transcription factor Pf-Rel, which belongs to the Rel/nuclear factor-κB family, was shown to regulate transcription at the Nacrein promoter in the pearl oyster Pinctada fucata. Here, we further explored the transcriptional regulation mechanisms of Pf-Rel on the matrix proteins Prismalin-14 and MSI60. The relative expression levels of Prismalin-14 and MSI60 were high in the mantle edge and mantle pallial tissues of P. fucata. These three genes were significantly up-regulated after shell notching, suggesting that they might play important roles during shell formation. Importantly, Pf-Rel gene knockdown by RNA interference led to down-regulation of Prismalin-14 and MSI60 expression. In transient co-transfection assays, Pf-Rel significantly up-regulated the promoter activities of the Prismalin-14 and MSI60 genes in a dose-dependent manner. Furthermore, the promoter regions of Prismalin-14 (-1794 to -1599bp) and MSI60 (-2244 to -1141bp) were required for the activation by Pf-Rel. Altogether, these results suggest that the transcription factor Pf-Rel can up-regulate the expression of the matrix protein genes Prismalin-14 and MSI60 during shell formation in P. fucata, which improves our understanding of transcription regulation at the molecular level during molluscan shell development.