Four differentially expressed cDNAs in Callinectes sapidus containing the Rebers–Riddiford consensus sequence

Abstract

Decapod crustaceans such as Callinectes sapidus, the blue crab, provide unique opportunities to study proteins involved in biomineralization. Subsequent to each molt, the previously deposited soft cuticle is calcified while the postecdysial layers are simultaneously deposited and mineralized. Though the majority of the exoskeleton hardens, morphologically similar cuticle at the joints, called arthrodial membrane, remains flexible. It seems reasonable that hypodermal cells producing these cuticle types should be synthesizing proteins that regulate mineralization. Data presented here are consistent with this hypothesis, showing that transcripts coding for proteins containing the chitin-binding Rebers–Riddiford (RR) consensus sequence (Gx 8Gx 7YxAxExGYx 7Px 2P) are differentially expressed. Two RR-containing transcripts, CsAMP8.1 and CsAMP6.0, are found only in arthrodial membrane and are expressed uniformly both before and after ecdysis. They have high sequence homology with RR-containing proteins from uncalcified portions of the cuticle of Cancer pagurus, Penaeus japonicus, and Homarus americanus. The other two transcripts, CsCP8.5 and CsCP8.2, are expressed solely in premolt and in hypodermis depositing calcifying cuticle rather than arthrodial membrane. They have high sequence homology with calcification-associated peptides containing the RR sequence obtained from the calcified cuticle of Procambarus clarkii. This suggests possible involvement in the postmolt mineralization of the pre-ecdysial cuticle.