Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System

Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정

  • 이수복 (부산대학교 자연과학대학 분자생물학과) ;
  • 김규원 (서울대학교 약학대학 생화학교실) ;
  • 배문경 (부산대학교 자연과학대학 분자생물학과) ;
  • 배명호 (부산대학교 자연과학대학 분자생물학과) ;
  • 정주원 (부산대학교 자연과학대학 분자생물학과) ;
  • 안미영 (부산대학교 자연과학대학 분자생물학과) ;
  • 김영진 (부산대학교 자연과학대학 분자생물학과)
  • Published : 2004.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

본 연구는 redox regulator로 알려 진 Ref-1 (Redox factor-1)과 결합하는 새로운 단백질을protein-protein interaction의 원리를 이용한 방법인 yeast two-hybrid assay로 검색, 동정하고, 검색된 단백질의 in vitro, in vivo 기능을 규명하는 데 그 목적을 두고, mouse 11-day Embryo cNA library를 prey로, full length REF-1을 bait로 하여 yeast strain 인 HF7C에 cotransformatiom시킨 후 histidine, leucine, tryptophan이 결핍된 SD plate에서 키워 자란 yeast transformants를 $\beta$-galactosidaseassay하여 screening하여 분리한 세 개의 clone중 한 clone이 DNA sequencing으로 확인한 결과 mouse thioredoxin임을 확인하였다.

Keywords

References

  1. Cancer Res. v.59 no.23 Egr-1 mediates transcriptional activation of IGF-Ⅱ gene in response to hypoxia Bae,S.K.;M.H.Bae;M.Y.Ahn;M.J.Son;Y.M.Lee;M.K.Bae;O.H.Lee;B.C.Park;K.W.Kim
  2. Mol. Cell. Biol. v.20 no.1 Redox-regulated recruitment of the transcriptional coactivators CREB-binding protein and SRC-1 to hypoxia-inducible factor 1alpha Carrero,P.;K.Okamoto;P.Coumailleau;S.O'Brien;H.Tanaka;L.Poellinger https://doi.org/10.1128/MCB.20.1.402-415.2000
  3. J. Exp. Med. v.192 no.6 Nitric oxide-dependent activation of p53 suppresses bleomycin-induced apoptosis in the lung Davis,D.W.;D.A.Weidner;A.Holian;D.J.McConkey https://doi.org/10.1084/jem.192.6.857
  4. Proc. Natl. Acad. Sci. U.S.A. v.88 Cloning and expression of APE, the cDNA encoding the major human apurinic endonuclease: definition of a family of DNA repair enzymes Demple,B.;T.Herman;D.S.Chen https://doi.org/10.1073/pnas.88.24.11450
  5. Curr. Opinion Cell Biol. v.10 Oxygen radicals and signaling Finkel,T. https://doi.org/10.1016/S0955-0674(98)80147-6
  6. Proc. Natl. Acad. Sci. USA v.94 AP-1 transciptional activity is regulated by a direct association between thioredoxin and REF-1 Hirota,K.;M.Matsui;S.Iwata;A.Nishiyama;K.Mori;J.Yodoi https://doi.org/10.1073/pnas.94.8.3633
  7. DNA Cell Biol. v.12 no.3 Characterization of the DNA-binding properties of the early growth response-1 (Egr-1) transciption factor: evidence for modulation by a redox mechanism Huang,R.P;E.D.Adamson https://doi.org/10.1089/dna.1993.12.265
  8. Science v.283 no.5398 The transcriptional program in the response of human fibroblasts to serum Iyer,V.R.;M.B.Eisen;E.T.Ross;G.Schuler;T.Moore;J.C.F.Lee;J.M.Trent;L.M.Staudt;J.Jr.Hudson;M.S.Boruski;D.Shalon;D.Lashkari;D.Botstein;P.O.Brown https://doi.org/10.1126/science.283.5398.83
  9. Genes Dev. v.11 Identification of redox/repair protein REF-1 as a potent activator of p53 Jayaraman,L.;K.G.Murthy;C.Zhu;T.Curran;S.Xanthoudakis;C.Prives https://doi.org/10.1101/gad.11.5.558
  10. Cell Signal. v.11 Redox regulation of cellular signaling Kamata,H.;H.Hirata https://doi.org/10.1016/S0898-6568(98)00037-0
  11. Cancer Res. v.58 no.2 Insulin-like growth factor Ⅱ induced by hypoxia may contribute to angiogenesis of human hepatocellular carcinoma Kim,K.W.;S.K.Bae;O.H.Lee;M.H.Bae;M.J.Lee;B.C.Park
  12. Biochem. Biophys. Res. Commun. v.264 no.3 Sphingosine 1-Phosphate Induces Anginogenesis: Its Anginogenic Action and Signaling Mechanism in Human Umbilical Vein Endithelial Cells Lee,O.H.;Y.M.Kim;Y.M.Lee;E.J.Moon;D.J.Lee;J.H.Kim;K.W.Kim;Y.G.Kwon https://doi.org/10.1006/bbrc.1999.1586
  13. Nat. Genet. v.17 Genetic interaction between HAPI/REF-1 and p53 Meira,L.B.;D.L.Cheo;R.E.Hammer;D.K.Burns;A.Reis;E.C.Friedberg https://doi.org/10.1038/ng1097-145
  14. Annu. Rev. Immunol. v.15 Redox regulation of cellular activation Nakamura,H.;K.Nakamura;J.Yodoi https://doi.org/10.1146/annurev.immunol.15.1.351
  15. Dev. Biol. v.199 no.1 The transition to endore-duplication in trophoblast gianr cells is reglulated by the mSNA zinc finger transcription factor Nakayama,H.;I.C.Scott;J.C.Cross https://doi.org/10.1006/dbio.1998.8914
  16. Cell v.88 no.2 Endostatin, an endongenous inhibitor of angiogenesis and tumor growth O'Reilly,M.S.;T.Boehm;Y.Shing;N.Fukai;G.Vasios;W.S.Lane;E.Flynn;J.R.Birkhead;B.R.Olsen;J.Folkman https://doi.org/10.1016/S0092-8674(00)81848-6
  17. Proc. Natl. Acad. Sci. U.S.A. v.96 no.16 Distinctive gene expression patterns in hyman mammary epithelial cells and breast cancers Perou,C.M.;S.S.Jeffrey;M.van,de,Rijn;C.A.Rees;M.B.Eisen;D.T.Ross;A.Pergamenschikov;C.F.Williams;S.X.Zhu;J.C.Lee;D.Lashkari;D.Shalon;P.O.Brown;D.Botstein https://doi.org/10.1073/pnas.96.16.9212
  18. Toxicology. v.147 no.1 Implications of oxidative stress and inflammatory process in the cytotoxicity of capsaicin in human endothelial cells: lack of DNA strand breakage Richeux,F.;M.Cascante;R.Ennamany;D.Sanchez;A.Sanni;D.Saboureau;E.E.Creppy https://doi.org/10.1016/S0300-483X(00)00184-0
  19. J. Biol. Chem. v.266 cDNA and deduced amino acid sequence of a mouse DNA repair enzyme(APEX nuclease) with signification homology to Escherichia coli exonuclease Ⅲ Seki,S.;K.Akiyama;S.Watanabe;M.Hatsushika;S.Ikeda;K.Tsutsui
  20. Curr. Opin. Genet. Dev. v.8 no.5 Hypoxia-inducible factor 1: master regulator of O₂homeostasis Semenza,G.L. https://doi.org/10.1016/S0959-437X(98)80016-6
  21. FASEB. v.10 Antioxidant and dedox regulation of gene transcription Sen.C.K.;L.Packer
  22. Biochem. Pharmacol. v.55 Redox signaling and the emerging therapeutic potential of thiol antioxidants Sen.C.K. https://doi.org/10.1016/S0006-2952(97)00672-2
  23. Curr. Top. Cell. Regul. v.36 Cellular thiols and redox-regulated signal transduction Sen.C.K.
  24. Proc. Natl. Acad. Sci. U.S.A. v.96 no.7 Overexpression of thioredoxin in transgenic mice attenuates focal ischemic brain damage Takagi,Y.;A.Mitsui;A.Nishiyama;K.Nozaki;H.Sono;Y.Gon;N.Hashimoto;J.Yodoi https://doi.org/10.1073/pnas.96.7.4131
  25. J. Biol. Chem. v.274 no.50 Thioredoxin-dependent redox regulation of p53-mediated p21 activation Ueno,M.;H.Masutani;R.J.Arai;A.Yamauchi;K.Hirota;T.Sakai;T.Inamoto;Y.Yamaoka;J.Yodoi;T.Nikaido https://doi.org/10.1074/jbc.274.50.35809
  26. J. Neurochem. v.73 no.6 Activation of AP-1 and nuclear factor-kappaB transcription factors is involved in hydrogen peroxide-induced apoptotic cell death of oligodendrocytes Vollgraf,U.;M.Wegner;C. Richter-Landsberg https://doi.org/10.1046/j.1471-4159.1999.0732501.x
  27. Nucleic Acids Res. v.22 no.23 A role for the human DNA repair enzyme HAP1 in cellular protection against DNA damaging agents and hypoxic stress Walker,L.J;R.B.Craig;A.L.Harris;I.D.Hickson https://doi.org/10.1093/nar/22.23.4884
  28. EMBO J. v.11 Identification and characterization of REF-1, a nuclear protein that facilitates AP-1 DNA-binding activity Xanthoudakis,S.;T.Curran
  29. EMBO J. v.11 no.9 Redox activation of Fos-Jun DNA binding activity is mediated by a DNA repair enzyme Xanthoudakis,S.;G.Miao;F.Wang;Y.C.E.Pan;T.Curran
  30. Proc. Natl. Acad. Sci. U.S.A. v.93 no.17 The redox/DNA repair protein,REF-1, is essential for early embryonic development in mice Xanthoudakis,S.;R.J.Smeyne;J.D.Wallace;Y.Curran https://doi.org/10.1073/pnas.93.17.8919
  31. Mol. Cell. Biol. v.14 no.9 Activation of AP-1 and of a nuclear redox factor, REF-1, in the response of HT29 colon cancer cells to hypoxia Yao,K.S.;S.Xanthoudakis;T.Curran;P.J.O'Dwyer https://doi.org/10.1128/MCB.14.9.5997
  32. Adv. Cancer Res. v.57 ADF, a growth-promoting factor derived from adult T cell leukemia and homologous to thioredoxin:onvolvement in lymphocyte immortalization by HTLV-I and EBV Yodoi,J.;T.Tursz https://doi.org/10.1016/S0065-230X(08)61004-0