Abstract
Kinase-family with sequence similarity 20, member C (Fam20C) is a protein kinase, which can phosphorylate biomineralization related proteins in vertebrate animals. However, the function of Fam20C in invertebrate animals especially the role in biomineralization is still unknown. Herein, we cloned the cDNA of fam20C from the pearl oyster, Pinctada fucata. It is showed that the expression of fam20C in the mantle edge was much higher than other tissues. In situ hybridization showed that fam20C was expressed mostly in the outer epithelial cells of the middle fold, indicating it may play important roles in the shell formation. Besides, fam20C expression increased greatly in the D-shape stage of pearl oyster development, when the shell was first formed. During the shell repair process, the expression level of fam20C increased 1.5 times at 6 h after shell notching. Knockdown of fam20C in vivo by RNA interference resulted in abnormally stacking of calcium carbonate crystals at the edges of nacre tablets, showing direct evidence that fam20C participates in the shell formation. This study provides an insight into the role of kinase protein in the shell formation in mollusk and broaden our understanding of biomineralization mechanism.
Highlights
Biomineralization is a process that minerals are formed by organisms[1]
A dentin-matrix protein-like (DMP-like), exhibiting a remarkable Fam20C domain was detected in two freshwater mussels unionoid proteomes24. cfMSP-1, an extremely acidic matrix protein involved in shell formation of the scallop Chlamys farreri, had eight S-D-E motifs, which were possibly phosphorylated by Fam20C25
The human and mouse fam20C gene were more complex than other species. fam20C of P. fucata, Lottia gigantean and Crassostrea gigas lacked transmembrane peptide, while all three vertebrates species and Villosa lienosa had transmembrane peptides
Summary
Biomineralization is a process that minerals are formed by organisms[1]. Pearl oyster, Pinctada fucata is one of the most important cultured pearl species in China, and is a good model species to study biomineralization[2]. Fam20C was highly expressed in dental tissues in mice and was highly expressed in odontoblast during the developing process of mice teeth, indicating that the Fam20C was closely related to mineral formation[17] These studies confirmed that Fam20C was closely involved in the biomineralization process of vertebrate animals. CfMSP-1, an extremely acidic matrix protein involved in shell formation of the scallop Chlamys farreri, had eight S-D-E motifs, which were possibly phosphorylated by Fam20C25. Despite these studies, no direct evidence has been given on the role of Fam20C in shell formation. Shell notching experiment and RNA interference were performed to investigate the role of Fam20C in biomineralization in vivo
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