Characterization of the shrimp eyestalk cDNA encoding a novel fushi tarazu-factor 1 (FTZ-F1)

Abstract

To study the role of ecdysone and the ecdysone inducible gene in the regulation of molting and development in crustaceans, we have cloned a cDNA encoding an orphan nuclear receptor family member from the eyestalk of the shrimp Metapenaeus ensis. The size of the cDNA is 4.3 kb with the longest open reading frame (ORF) encoding a protein of 545 amino acid residues. The deduced amino acid sequence of the shrimp cDNA consists of regions that are characteristic of those of the nuclear hormone receptors. It shows a high degree of amino acid sequence identity in the DNA binding domain, ligand binding domain and the FTZ box as compared to those of invertebrates and vertebrates. Unlike the insects Drosophila melanogaster and Bombyx mori, an AF2 transactivation domain was present in the shrimp FTZ-F1. Northern blot analysis using total RNA indicated that the FTZ-F1 mRNA could also be detected in the mature ovary. Northern blot analysis and RT-PCR analysis showed that the shrimp FTZ-F1 transcripts could be detected in the ovary, newly hatched nauplius, testis, eyestalk and epidermis of the adult shrimp. Although the cDNA clone was isolated from the eyestalk library, the shrimp FTZ-F1 appeared to express most abundantly in the mature oocytes. The presence of abundant FTZ-F1 specific maternal message in the late vitellogenic ovary and early nauplius indicates that it may be important for the early embryonic and larval development of the shrimp. Interestingly, shrimp FTZ-F1 can also be found in testis of the male shrimp. The presence of FTZ-F1 in other tissues such as epidermis suggests that it may also be involved in other physiological processes such as molting.