Cloning and expression of ecdysone receptor (EcR) from the intertidal copepod, Tigriopus japonicus.
Ecdysteroids are steroid hormones that play an important role in development, growth, molting of larva, and reproduction in the Arthropoda. The effect of ecdysteroids is mediated by its binding to ecdysteroid receptor (EcR). To investigate the role of EcR during development and the effect to environmental stressors on EcR expression in a copepod, we isolated and characterized cDNA and 5'-promoter region of the Tigriopus japonicus EcR (TJ-EcR), and studied mRNA expression pattern. The full-length TJ-EcR cDNA sequence was 1962bp in length and the open reading frame encoded 546 amino acids. The deduced TJ-EcR protein contained well-conserved DNA-binding domain and ligand-binding domain. Phylogenetic analysis revealed that TJ-EcR was clustered with the EcR of other crustaceans. TJ-EcR mRNA was expressed in a developmental stage-specific manner: high in early developmental stages and low in the adult stage. Significantly elevated expression of the TJ-EcR gene in adults was detected at hypersalinity (42ppt) and high temperature (35 degrees C) condition. The 5'-flanking region of TJ-EcR gene contains heat shock protein 70 response elements, implying that the environmental stressors may affect its expression via the stress-sensor. In addition, bisphenol A (100microg/L) repressed TJ-EcR expression. Our results suggest that TJ-EcR could be a biomarker for the monitoring of the impact of environmental stressors in copepods.
Hwang
Dae-Sik
DS
Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University, Seoul 133-791, South Korea.
Lee
Jin-Seon
JS
Lee
Kyun-Woo
KW
Rhee
Jae-Sung
JS
Han
Jeonghoon
J
Lee
Jehee
J
Park
Gyung Soo
GS
Lee
Young-Mi
YM
Lee
Jae-Seong
JS
D024506:5' Flanking Region / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001559:Benzhydryl Compounds; D003001:Cloning, Molecular; D033342:Copepoda / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D052244:Endocrine Disruptors / Q000633:toxicity; D015870:Gene Expression / Q000187:drug effects; D008024:Ligands; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010636:Phenols / Q000633:toxicity; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D054712:Salinity; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013312:Stress, Physiological; D000098826:Bisphenol A Compounds
D001559:Benzhydryl Compounds; D018076:DNA, Complementary; D052244:Endocrine Disruptors; D008024:Ligands; D010636:Phenols; D012333:RNA, Messenger; D011987:Receptors, Steroid; C034808:ecdysteroid receptor; C006780:bisphenol A; D000098826:Bisphenol A Compounds