Molecular cloning and expression analysis of ecdysone receptor and retinoid X receptor from the kuruma prawn, Marsupenaeus japonicus.
Two cDNAs encoding EcR (MjEcR) and RXR (MjRXR) were cloned and sequenced from the kuruma prawn Marsupenaeus japonicus using PCR techniques. The amino acid sequence of MjEcR was similar to that of known EcR especially in the ligand binding domain (LBD) of insect EcR. The DNA binding domain of MjRXR showed higher homology with that of insect USP (>90% identity) than vertebrate RXR ( approximately 85% identity), while LBD of MjRXR is more homologous with that of vertebrate RXR ( approximately 65% identity) than that of insect USP (30-60% identity). The transcripts of MjEcR and MjRXR were expressed in all tissues examined and in particular, highly in Y-organ and heart and in ovary and heart, respectively. Quantitative real-time PCR analyses revealed that the expression level of MjEcR in hepatopancreas and thoracic muscle increased from intermolt to premolt stages. The analyses also showed that the expressions of MjEcR and MjRXR were regulated in a tissue-specific manner. No significant changes were observed in reproductive organs throughout the molting stages, and MjRXR was expressed much more than MjEcR at all stages. These data suggest that MjRXR mediates a certain hormonal signal related to reproduction.
Asazuma
Hideaki
H
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo, Tokyo 113-8657, Japan.
Nagata
Shinji
S
Kono
Michiko
M
Nagasawa
Hiromichi
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D018978:Molting / Q000235:genetics; D009928:Organ Specificity; D033561:Penaeidae / Q000235:genetics*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D047488:Retinoid X Receptors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D012333:RNA, Messenger; D011987:Receptors, Steroid; D047488:Retinoid X Receptors; C055469:ecdysone receptor