Transcriptional Activity of an Estrogen Receptor β Subtype in the Medaka Oryzias dancena
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In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, foldactivation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.
Abstract
In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, foldactivation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.
참고문헌 (27)
- Antal MC, "Estrogen dependent activation function of ERβ is essential for the sexual behavior of mouse females", Proc Natl Acad Sci USA, 109, pp.19822~19827 (2012)
- Chakraborty T, "Differential expression of three estrogen receptor subtype mRNAs in gonads and liver from embryos to adults of the medaka, Oryzias latipes", Mol Cell Endocrinol, 333, pp.47~54 (2011)
- Cho YS, "Functional ability of cytoskeletal β-actin regulator to drive constitutive and ubiquitous expression of a fluorescent reporter throughout the life cycle of transgenic marine medaka Oryzias dancena", Transgenic Res, 20, pp.1333~1355 (2011)
- Cho YS, "Characterization of estrogen-responsive transgenic marine medaka Oryzias dancena germlines harboring red fluorescent protein gene under the control by endogenous choriogenin H promoter", Transgenic Res, 22, pp.501~517 (2013)
- Evans RM, "The steroid and thyroid hormone receptor superfamily", Science, 240, pp.889~895 (1988)
- Glass CK, "The coregulator exchange in transcriptional functions of nuclear receptors", Genes Dev, 14, pp.121~141 (2000)
- Haldosén LA, "Estrogen receptor beta in breast cancer", Mol Cell Endocrinol, 382, pp.665~672 (2014)
- Jones DT, "The rapid generation of mutation data matrices from protein sequences", Comput Appl Biosci, 8, pp.275~282 (1992)
- Jung DJ, "Agonist-dependent repression mediated by mutant estrogen receptor alpha that lacks the activation function 2 core domain", J Biol Chem, 276, pp.37280~37283 (2001)
- Kayo D, "Gene knockout analysis reveals essentiality of estrogen receptor β1 (Esr2a) for female reproduction in medaka", Sci Rep, 9, pp.8868~ (2019)
- 김현규, "Gene Expression of Aromatases, Steroid Receptor, GnRH and GTHs in the Brain during the Formation of Ovarian Cavity in Red Spotted Grouper, Epinephelus akaara", 발생과 생식, 20(4), pp.356~377 (2016), 한국발생생물학회
- Lange IG, "Evolution of oestrogen functions in vertebrates", J Steroid Biochem Mol Biol, 83, pp.219~226 (2002)
- Lee Pow CSD, "Sharing the roles: An assessment of Japanese medaka estrogen receptors in vitellogenin induction", Environ Sci Technol, 50, pp.8886~8895 (2016)
- Maeng S, "Expression of gonadotropin subunit genes following 4-nonylphenol exposure in masu salmon: Effects on transcript levels and promoter activities via estrogen receptor alpha", Comp Biochem Physiol B Biochem Mol Biol, 142, pp.383~390 (2005)
- Mu WJ, "Molecular cloning and expression analysis of estrogen receptor betas (ERβ1 and ERβ2) during gonad development in the Korean rockfish, Sebastes schlegeli", Gene, 523, pp.39~49 (2013)
- Nagler JJ, "The complete nuclear estrogen receptor family in the rainbow trout: Discovery of the novel ERalpha2 and both ERbeta isoforms", Gene, 392, pp.164~173 (2007)
- Nagler JJ, "Estrogen receptor mRNA expression patterns in the liver and ovary of female rainbow trout over a complete reproductive cycle", Gen Comp Endocrinol, 178, pp.556~561 (2012)
- Nelson ER, "Estrogen receptor function and regulation in fish and other vertebrates", Gen Comp Endocrinol, 192, pp.15~24 (2013)
- Ogawa S, "The complete primary structure of human estrogen receptor beta (hER beta)and its heterodimerization with ER alpha in vivo and in vitro", Biochem Biophys Res Commun, 243, pp.122~126 (1998)
- Rasier G, "Female sexual maturation and reproduction after prepubertal exposure to estrogens and endocrine disrupting chemicals: A review of rodent and human data", Mol Cell Endocrinol, 254-255, pp.187~201 (2006)
- Sabo-Attwood T, "Differential expression of largemouth bass (Micropterus salmoides) estrogen receptor isotypes alpha, beta, and gamma by estradiol", Mol Cell Endocrinol, 218, pp.107~118 (2004)
- Schmittgen TD, "Analyzing real-time PCR data by the comparative C(T) method", Nat Protoc, 3, pp.1101~1108 (2008)
- Tamura K, "MEGA6: Molecular evolutionary genetics analysis version 6.0", Mol Biol Evol, 30, pp.2725~2729 (2013)
- Thomas C, "The different roles of ER subtypes in cancer biology and therapy", Nat Rev Cancer, 11, pp.597~608 (2011)
- Webb P, "ERbeta binds N-CoR in the presence of estrogens via an LXXLL-like motif in the N-CoR C-terminus", Nucl Recept, 1, pp.4~ (2003)
- Zavacki AM, "Activation of the orphan receptor RIP14 by retinoids", Proc Natl Acad Sci USA, 94, pp.7909~7914 (1997)
- Zhao C, "Estrogen receptor beta: An overview and update", Nucl Recept Signal, 6, pp.e003~ (2008)