둥근성게(Strongylocentrotus nudus) Estrogen Receptor-Related Receptor(ERR)의 초기 발생시 유전자 발현 패턴과 전사 활성

Gene Expression Pattern during Early Embryogenesis and Transcriptional Activities of Estrogen Receptor-Related Receptor(ERR) in Sea Urchin, Strongylocentrotus nudus

  • 맹세정 (강릉원주대학교 해양분자생명공학) ;
  • 김미순 (강릉원주대학교 해양분자생명공학) ;
  • 손영창 (강릉원주대학교 해양분자생명공학)
  • Maeng, Se-Jung (Dept. of Marine Molecular Biotechnology, Gangneung-Wonju National University) ;
  • Kim, Mi-Soon (Dept. of Marine Molecular Biotechnology, Gangneung-Wonju National University) ;
  • Sohn, Young-Chang (Dept. of Marine Molecular Biotechnology, Gangneung-Wonju National University)
  • 발행 : 2009.12.31

초록

구조적으로 estrogen 수용체(estrogen receptor, ER)와 유사한 estrogen receptor-related receptor(ERR)는 포유동물에서 배발생 후기에 외배엽 형성과 관련되어 있다고 알려진 고아핵수용체(orphan nuclear receptor)이다. ERR은 ER과 DNA binding domain의 보존성은 유사하지만, ligand 결합 및 전사 활성은 다르다. 포유동물의 ERR에 관한 연구에 비하여 해양 무척추동물의 ERR에 대한 기능 연구는 매우 부족하다. 본 연구에서는 우리나라 동해안에 주로 서식하는 둥근성게(Strongylocentrotus nudus) ERR의 초기 발생기 유전자 발현 변화와 전사 활성 기능을 조사하기 위해 먼저 polymerase chain reaction(PCR)을 이용하여 cDNA를 동정하였다. 둥근성게 ERR은 S. purpuratus와 91%의 높은 상동성을 보였으며, 계통수 분석을 통해 무척추동물 ERR의 clade에 포함되는 것을 알았다. 둥근성게 배발생 시기에 ERR 유전자 발현을 알아보기 위하여 real-time PCR을 실시한 결과, 4~64세포기와 유생기에 mRNA level이 높은 경향을 나타내었다. 또한 호르몬 및 co-regulator에 의한 둥근성게 ERR의 전사 활성을 조사한 결과, 호르몬에 의한 특이성은 확인되지 않았지만, peroxisome proliferator activated receptor-(PPAR) $\gamma$ coactivator $1\alpha$(PGC-$1\alpha$)가 둥근성게 ERR의 coactivator임을 입증할 수 있었다. 이 연구 결과는 향후 새로운 ligand 발굴과 coregulator와 의 상호작용 연구에 도움이 될 것으로 사료된다.

The estrogen receptor-related receptors (ERRs) are a group of nuclear receptor superfamily of transcription factors. ERRs and estrogen receptors (ERs) have overlapping affinities for coactivators and DNA binding sites, but differ markedly in ligand binding and activation. The three mammalian ERR genes have been implicated in diverse physiological processes ranging from placental development to maintenance of bone density, whereas the molecular diversity, function, and regulation of ERRs in non-mammalian species are not well understood. In the present study, to investigate the involvement of ERR in transcription and embryogenesis in marine invertebrates, a cDNA encoding ERR (SnERR) was cloned from the gonad in Strongylocentrotus nudus, by polymerase chain reaction (PCR). The amino acid sequence of SnERR showed high homology with that of S. purpuratus (91%). A phylogenetic tree clearly showed that SnERR is a member of the ERR family and clustered in echinodermata group as supported by a high bootstrap value. We examined gene expression of SnERR during embryonic development of S. nudus using real-time PCR. During the embryonic development, the mRNA of ERR was significantly high levels in early development stages (4~64 cell) and larval stages. The SnERR slightly activated transcription through the classical estrogen response elements (EREs) in the presence of genistein. In addition, peroxisome proliferator-activated receptor $\gamma$ coactivator (PGC)-$1\alpha$ knwon as a coactivator of ERR enhanced the snERR-mediated transactivation, suggesting that the PGC-$1\alpha$ is a coactivator of SnERR.

키워드

참고문헌

  1. Ao A, Wang H, Kamarajugadda S, Lu J (2008) Involvement of estrogen-related receptors in transcriptional response to hypoxia and growth of solid tumors. Proc Natl Acad Sci USA 3;105:7821-7826. https://doi.org/10.1073/pnas.0711677105
  2. Bardet PL, Obrecht-Pflumio S, Thisse C, Laudet V, Thisse B, Vanacker JM (2004) Cloning and developmental expression of five estrogen-receptor related genes in the zebrafish. Dev Genes Evol 214:240-249. https://doi.org/10.1007/s00427-004-0404-6
  3. Bardet PL, Schubert M, Horard B, Holland LZ, Laudet V, Holland ND, Vanacker JM (2005) Expression of estrogenreceptor related receptors in amphioxus and zebrafish: implications for the evolution of posterior brain segmentation at the invertebrate-to-vertebrate transition. Evol Dev 7:223-233. https://doi.org/10.1111/j.1525-142X.2005.05025.x
  4. Bonnelye E, Vanacker JM, Spruyt N, Alric S, Fournier B, Desbiens X, Laudet V (1997) Expression of the estrogenrelated receptor 1 (ERR-1) orphan receptor during mouse development. Mech Dev 65:71-85. https://doi.org/10.1016/S0925-4773(97)00059-2
  5. Chen F, Zhang Q, McDonald T, Davidoff M, Bailey W, Bai C, Liu Q, Caskey C (1999) Identification of two hERR2-related novel nuclear receptors utlizing bioinformatics and inverse PCR. Gene 228:101-109. https://doi.org/10.1016/S0378-1119(98)00619-2
  6. Coward P, Lee D, Hull MV, Lehmann JM (2001) 4-Hydroxytamoxifen binds to and deactivates the estrogenrelated receptor ${\gamma}$. Proc Natl Acad Sci USA 98:8880- 8884. https://doi.org/10.1073/pnas.151244398
  7. Devine C, Hinman VF, Dengnan BM (2002) Evolution and developmental expression of nuclear receptor genes in the ascidian Herdmania. Dev Biol 46:687-692.
  8. Giguere V, Yang N, Segui P, Evans R (1988) Identification of a new class of steroid hormone receptors. Nature 331:91-94. https://doi.org/10.1038/331091a0
  9. Giguere V (2002) To ERR in the estrogen pathway. Trends Endocrinol Metab 13:220-225. https://doi.org/10.1016/S1043-2760(02)00592-1
  10. He C, Wang C, Li B, Xie F, Chen Y, Zuo Z (2009) Tissuespecific and embryonic expression of the retinoid X receptors in Sebastiscus marmoratus. Comp Biochem Physiol B Biochem Mol Biol 154:221-228. https://doi.org/10.1016/j.cbpb.2009.06.006
  11. Heard DJ, Norby P, Holloway J, Vissing H (2000) Human ERR${\gamma}$, a third member of the estrogen receptor-related receptor (ERR) subfamily of orphan nuclear receptors: tissue-specific isoforms are expressed during development and in the adult. Mol Endocrinol 14:382-392. https://doi.org/10.1210/me.14.3.382
  12. Hong H, Yang L, Stallcup MR (1999) Hormone-independent transcriptional activation and coactivator binding by novel orphan receptor ERR3. J Biol Chem 274:22618- 22626. https://doi.org/10.1074/jbc.274.32.22618
  13. Hood L, Davidson EH, Cameron RA, Ettensohn C, Wray G (2001) The Sea Urchin Genome Project-Introduction. http://sugp.caltech.edu/intro/ (March 10, 2003).
  14. Horard B, Castet A, Bardet PL, Laudet V, Cavailles V, Vanacker JM (2004) Dimerization is required for transactivation by estrogen-receptor-related (ERR) orphan receptors: evidence from amphioxus ERR. J Mol Endocrinol 33:493-509. https://doi.org/10.1677/jme.1.01538
  15. Howard-Ashby M, Materna SC, Brown CT, Tu Q, Oliveri P, Cameron RA, Davidson EH (2006a) Gene families encoding transcription factors expressed in early development of Strongylocentrotus purpuratus. Dev Biol 300:90-107. https://doi.org/10.1016/j.ydbio.2006.08.033
  16. Howard-Ashby M, Materna SC, Brown CT, Tu Q, Oliveri P, Cameron RA, Davidson EH (2006b) High regulatory gene use in sea urchin embryogenesis: Implications for bilaterian development and evolution. Dev Biol 300: 27-34. https://doi.org/10.1016/j.ydbio.2006.10.016
  17. Jones BB, Ohno CK, Allenby G, Boffa MB, Levin AA, Grippo JF, Petkovich M (1995) New retinoid X receptor subtypes in zebra fish (Danio rerio) differentially modulate transcription and do not bind 9-cis retinoic acid. Mol Cell Biol 15:5226-5234.
  18. Jung Y, Sohn YC (2007) Gene expression of Smad3 and estrogen receptor-related receptor ${\beta}$ like 1 in sea urchin, Strongylocentrotus nudus. Dev Reprod 11:43-47.
  19. Jung Y, Maeng S, Sohn YC (2007) Effects of estradiol-17 ${\beta}$ and nonylphenol on mRNA expression of estrogen receptor-related receptor ${\beta}$ like 1 and early embryogenesis in sea urchin, Strongylocentrotus nudus. Dev Reprod 11:179-185.
  20. Liu D, Zhang Z, Gladwell W, Teng CT (2003) Estrogen stimulates estrogen-related receptor alpha gene expression through conserved hormone response elements. Endocrinology 144:4894-4904. https://doi.org/10.1210/en.2003-0432
  21. Luo J, Sladek R, Bader JA, Matthyssen A, Rossant J, Giguere V (1997) Placental abnormalities in mouse embryos lacking the orphan nuclear receptor ERR-beta. Nature 388:778-782. https://doi.org/10.1038/42022
  22. Maeng S, Jung Y, Choi E, Jeon JK, Kim S, Gen K, Sohn YC (2005) 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 142:383-390. https://doi.org/10.1016/j.cbpb.2005.08.009
  23. Mitsunaga K, Araki K, Mizusaki J, Morohashi KI, Haruna K, Nakagata N, Giguere V, Yamamura KI, Abe K (2004) Loss of PGC-specific expression of the orphan nuclear receptor ERR-results in reduction of germ cell number in mouse embryos. Mech Dev 121:237-246. https://doi.org/10.1016/j.mod.2004.01.006
  24. Park W, Kim GJ, Choi HS, Vanacker JM, Sohn YC (2009) Conserved properties of a urochordate estrogen receptorrelated receptor (ERR) with mammalian ERR alpha. Biochim Biophys Acta 1789:125-134. https://doi.org/10.1016/j.bbagrm.2008.08.011
  25. Pettersson K, Svensson K, Mattsson R, Carlsson B, Ohlsson R, Berkenstam A (1996) Expression of a novel member of estrogen response element-binding nuclear receptors is restricted to the early stages of chorion formation during mouse embryogenesis. Mech Dev 54:211-223. https://doi.org/10.1016/0925-4773(95)00479-3
  26. Tarrant AM, Greytak SR, Callard GV, Hahn ME (2006) Estrogen receptor-related receptors in the killifish Fundulus heteroclitus: diversity, expression, and estrogen responsiveness. J Mol Endocrinol 37:105-120. https://doi.org/10.1677/jme.1.01976
  27. Tremblay G, Bergeron D, Giguere V (2001a) 4-hydroxytamoxifen is an isoform-specific inhibitor of orphan estrogen-receptorrelated (ERR) nuclear receptors ${\beta}$ and ${\gamma}$. Endocrinology 142:4572-4575. https://doi.org/10.1210/en.142.10.4572
  28. Tremblay G, Kunath T, Bergeron D, Lapointe L, Champigny C, Bader JA, Rossant J, Giguere V (2001b) Diethystilbestrol regulates trophoblast stem cell differentiation as a ligand of orphan nuclear receptor ERR${\beta}$. Genes Dev 15:833- 835. https://doi.org/10.1101/gad.873401
  29. Vanacker JM, Pettersson K, Gustafsson JA, Laudet V (1999a) Transcriptional targets shared by estrogen receptorrelated receptors (ERRs) and estrogen receptor (ER)${\alpha}$, but not by ER${\beta}$. EMBO J 18:4270-4279. https://doi.org/10.1093/emboj/18.15.4270
  30. Vanacker JM, Pettersson K, Gustafsson JA, Laudet V (1999b) Transcriptional activities of the orphan nuclear receptor ERR alpha (estrogen receptor-related receptoralpha). Mol Endocrinol 13:764-773. https://doi.org/10.1210/me.13.5.764
  31. Zavacki AM, Lehmann JM, Seol W, Willson TM, Kliewer SA, Moore DD (1997) Activation of the orphan receptor RIP14 byretinoids. Proc Natl Acad Sci USA 94:7909- 7914. https://doi.org/10.1073/pnas.94.15.7909
  32. Zhang ZB, Hu JY, Sai SX, Zhao YB, Huang C, Tian XJ (2008) Gene cloning, sequence analysis and tissue expression of estrogen-related receptor alpha (Erralpha) in Japanese medaka and its transcriptional responses after differential EDCs exposure. Huan Jing Ke Xue 29:3153-3158.