Identification of a gene encoding microphthalmia-associated transcription factor and its association with shell color in the clam Meretrix petechialis

The microphthalmia-associated transcription factor (MITF) is a master regulator of melanocyte development through the direct transcriptional control of related genes, e.g., the phenoloxidase gene. In this study, an MITF gene, MpMITF2, was identified in the clam Meretrix petechialis. The full-length cDNA of MpMITF2 was 2026 bp, and the molecular mass of the predicted protein was 42.6 kDa. A basic helix-loop-helix leucine zipper domain was detected in the deduced protein sequence, which can bind the E-box motif within the promoter of the downstream genes. The mRNA of MpMITF2 was more highly expressed in the mantle compared to the other four tissues. Furthermore, there was a significant difference in the mRNA expression of MpMITF2 among three clam strains with different shell colors. The protein level of MpMITF2 was also different among these strains. These results implied that MpMITF2 was associated with shell color formation in the clam M. petechialis. When the mRNA expression of MpMITF2 was knocked down, the new shell showed discontinuous pigment distribution, suggesting that the reduced expression of MpMITF2 influenced pigment synthesis. A gene encoding phenoloxidase (MpPO) was identified as related to the shell color of the clam and was also a putative downstream gene of MITF. Both the mRNA and protein levels of MpPO decreased significantly at 12 h post-MpMITF-suppression, suggesting that MpMITF2 is required for the expression of MpPO. Our results indicate the close relationships among MpMITF2, MpPO and shell color. This study implicates the role of MITF in shell color formation in the clam M. petechialis.