생명과학회지 (Journal of Life Science)

  • 제13권5호
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  • Pages.596-603
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  • 2003
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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누에 배양세포(Bm5)로부터 분리한 새로운 전사제어인자 ATFC의 특성분석

Isolation and Characterization of a Novel Transcription Factor ATFC Activated by ER Stress from Bombyx mori Bm5 Cell Lines

  • 구태원 (농촌진흥청 농업과학기술원) ;
  • 윤은영 (농촌진흥청 농업과학기술원) ;
  • 김성완 (농촌진흥청 농업과학기술원) ;
  • 최광호 (농촌진흥청 농업과학기술원) ;
  • 황재삼 (농촌진흥청 농업과학기술원) ;
  • 박수정 (충남대학교 의과대학) ;
  • 권오유 (충남대학교 의과대학) ;
  • 강석우 (농촌진흥청 농업과학기술원)
  • 발행 : 2003.10.01
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초록

누에배양세포주(Bm5)에 N-glycosylation 저해제인 tunicamycin를 처리하여 인위적으로 UPR를 유도하고 이로부터 cDNA 유전자은행을 제작한 후, 정상세포주에 비하여 발현량이 증가하는 40개의 차별화 발현 cDNA 클론을 선발하였다. 차별화발현 클론 중에서 기존에 밝혀진 전사제어인자와 1차 아미노산의 구조적 유사성을 나타내는 클론(ATFC)에 대하여 유전자의 구조와 발현특성을 분석한 결과는 다음과 같다. 유전자의 구조분석 결과, ATFC는 효모의 전사제어인자 Hac1p와 구조적으로 매우 유사하게 $\alpha$-helix 상의 7개 아미노산 잔기 마다 leucine이 7회 반복하여 출현하는 leucine zipper 모티프가 존재하고 있었으며, leucine zipper 바로 앞쪽에는 분자 샤페론이나 folding enzyme의 프로모터 부위에 존재하는 UPRE에 결합할 것으로 추정되는 염기성 아미노산이 풍부한 basic region이 존재하고 있었다. 또한, ATFC 유전자에 대하여 분자샤페론 및 folding enzyme의 전사 촉진 기능을 해석하기 위하여 누에 배양세포주(Bm5)에 각종 스트레스 유도제를 처리한 후 ATFC의 발현특성을 분석한 결과, 정상 세포주에서는 발현이 되지 않았으나 스트레스 유도제가 처리된 세포주에서는 ATFC의 전사체가 강하게 발현됨을 확인 할 수 있었다. 따라서 ATFC 유전자는 소포체 내의 정확하게 접혀지지 않았거나 조립되지 못한 단백질을 정확하게 접혀지게 하고 조립되게 하여 정상구조를 가지는 단백질로 재생하는 분자샤페론이나 folding enzyme의 전사를 촉진시키는 효모 Hac1p와 매우 유사한 기능을 수행할 것으로 추정할 수 있었다. 이상의 결과는, 효모를 제외한 모든 생물종에서 UPR pathway에 관련한 전사제어인자의 최초의 보고이다. 억제로 야기되는 것 같다.증진시키기 위해 행동변화단계에 따른 맞춤형 교육 프로그램을 개발하여 적용하였고, 그 결과, 행동변화단계별 교육 프로그램이 자궁경부암 조기검진의 수검 행동을 증진시키는데 효과적인 것으로 나타났다.lomus thermophilum, Thermotoga neapolitana 등에서 밝혀진 바와 같이 glutamic acid 부위가 xylanase의 활성부위라 여겨진다.倍), 수층(水層)이 약(約) 100배(100倍)로 나타나 홍삼(紅蔘)엑기스의 갈변색소형성(褐變色素形成)은 비효소적(非酵素的) 갈변반응(褐變反應)인 amino-carbonyl 반응(反應)이 주도적(主導的) 역할(役割)을 하고 있음을 알 수 있다. (6) 총당(總糖)과 갈변반응속도(褐變反應速度)는 유의성(有意性)이 있었으며 $100^{\circ}C$의 경우 20시간(20時間)에 가장 색도(色度)가 높아 갈변반응속도(褐變反應速度)가 0.2로 나타났다. the esophageal mucous cells pf Bryzoichthys lysimus contained small amount of neutral mucin, while on the other hand a feww mucous cells contained small amount of neutral mucin and minimal amount of sialomucin. But the esophageal mucous cells of Takifugu pardalis contained considerable amount of neutral mucin only.분해가 더욱 촉진되었으며, 30℃에서 교반 처리를 행한 경우가 10℃에서 교반 처리를 행한 경우 보다 지방분해가 더욱 촉진되었다. 산양유 원유는 30℃에서 교반 처리 시간이 연장되어도 지방분해는 뚜렷한 증가를 나타내지 않았다.와

Cells respond to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) by increasing transcription of genes encoding molecular chaperones and folding enzymes. The information is transmitted from the ER lumen to the nucleus by intracellular signaling pathway, called the unfolded protein response (UPR). To obtain genes related to UPR from B. mori, the cDNA library was constructed with mRNA isolated from Bm5 cell lines in which N-glycosylation was inhibited by tunicamycin treatment. From the cDNA library, we selected 40 clones that differentially expressed when cells were treated with tunicamycin. Among these clones, we have isolated ATFC gene showing similarity with Hac1p, encoding a bZIP transcription factor of 5. cerevisiae. Basic-leucine zipper (bZIP) domain in amino acid sequences of ATFC shared homology with yeast Hac1p. Also, ATFC is up-regulated by accumulation of unfolded proteins in the ER through the treatment of ER stress drugs. Therefore we suggest that ATFC represents a major component of the putative transcription factor responsible for the UPR leading to the induction of ER-localized stress proteins.

키워드

참고문헌

  1. Annu. Rev. Cell Dev. Biol. v.14 Intracellular signaling from the endoplasmic reticulum to the nucleus Chapman,R.;C.Sidrauski;P.Walter https://doi.org/10.1146/annurev.cellbio.14.1.459
  2. Cell. v.87 A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response Cox,J.S.;P.Walter https://doi.org/10.1016/S0092-8674(00)81360-4
  3. Trends Biochem. Sci. v.6 The tunicamycins-ueful tools for studies on glycoproteins Elbein,A.D. https://doi.org/10.1016/0968-0004(81)90080-3
  4. Nature v.328 Proteins as molecular chaperones Ellis,R.J. https://doi.org/10.1038/328378a0
  5. Phil. Trans Roy. Soc. Lnd. B. v.339 The general concept of molecular chaperones Ellis,R.J. https://doi.org/10.1098/rstb.1993.0023
  6. Nature v.355 Protein folding in the cell Gething,M.J.;J.Sambrook https://doi.org/10.1038/355033a0
  7. Kor. J. Genetics. v.23 Isolation and characterization of a gene encoding a protein disulfide isomerase from Bombyx mori Bm5 cell line Goo,T.W.;E.Y.Yun;J.S.Hwang;S.W.Kang;K.S.Lee;O.Y.Kwon
  8. J.Cell Biol. v.126 Quality control in the secretory pathway: retention of a misfolded viral membrane glycoprotein involves cycling between the ER, intermediate compartment, and Golgi apparatus Hammond,C.;A.Helenius https://doi.org/10.1083/jcb.126.1.41
  9. Biochem. J. v.355 Identification of the G13 (cAMP-response-element-binding protein-related protein) gene product related to activating transciption factor 6 as a transcriptional activator of the mammalian unfolded protein response Haze,K.;T.Okada;H.Yoshida;H.Yanagi;T.Yura;M.Negishi;K.Mori https://doi.org/10.1042/0264-6021:3550019
  10. Mol. Biol. Cell. v.10 Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress Haze,K.;H.Yoshida;H.Yanagi;T.Yura;K.Mori https://doi.org/10.1091/mbc.10.11.3787
  11. Ann. Res. Cell Biol. v.5 Protein oli-gomerization in the ER Hurtley,S.M;A.Helenius https://doi.org/10.1146/annurev.cb.05.110189.001425
  12. Nucleic Acids Res. v.19 Isolation of a lage number of novel mammalian genes by a differential cDNA library screening strategy HOOg https://doi.org/10.1093/nar/19.22.6123
  13. Nat. Cell Biol. v.3 Translational control by the ER transmembrane kinase/ribonuclease IRE1 under ER stress Iwawaki,T.;A.Hosoda;T.Okuda;Y.Kamigori;C.Nomura-Furuwatari;Y.Kimata;A.Tsuru;K.Kohno https://doi.org/10.1038/35055065
  14. Genes Dev. v.13 Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls Kaufman,R.J. https://doi.org/10.1101/gad.13.10.1211
  15. Proc. Natl. Acad. Sci. USA v.92 Molecular chaperones involved in protein degadation in the endoplasmic reticulum: quantitative interaction of the heat shock cognate protein BiP with partially folded immunoglobulin light chains that are degraded in the endoplasmic reticulum Knittler,M.R.;S.Dirks;I.G.Haas https://doi.org/10.1073/pnas.92.5.1764
  16. Mol. Cell. Biol. v.13 The promoter region of the yeast KAR2 (BiP) gene contains a regulatory domain that responds to the presence of unfolded protein in the endoplasmic reticulum Kohno,K.;K.Normington;J.F.Sambrook;M.J.Gething;K.Mori
  17. Curr. Opinion in Cell Biol. v.4 Mammalian stress response: induction of the glucose-regulated protein family Lee,A.S.
  18. J. Cell Biol. v.117 Misfolding and aggregation of newly synthesized proteins in the endoplasmic reticulum Marquardt,T.;A.Helenius https://doi.org/10.1083/jcb.117.3.505
  19. Eur. J. Biochem. v.251 A stress-inducible rat liver endoplasmic reticulum protein, ERp29 Mkrtchian,S.;C.Fang;U.Hellman;M.Ingelman-Sundberg https://doi.org/10.1046/j.1432-1327.1998.2510304.x
  20. Cell v.101 Tripartite management of unfolded proteins in the endoplasmic reticulum Mori,K. https://doi.org/10.1016/S0092-8674(00)80855-7
  21. Genes Cells v.1 Signaling from endoplasmic reticulum to the nucleus: transcription factor with a basic-leucine zipper motif is required for the unfolded protein-response pathway Mori,K.;T.Kawahara;H.Yoshida;H.Yanagi;T.Yura https://doi.org/10.1046/j.1365-2443.1996.d01-274.x
  22. J. Biol. Chem. v.273 Palindrome with spacer of one nucleotide is characteristic of the cis-acting unfolded protein response element in Saccharomyces cerevisiae Mori,K.;N.Ogawa;T.Kawahara;H.Yanagi;T.Yura https://doi.org/10.1074/jbc.273.16.9912
  23. EMBO J. v.11 A 22bp cis-acting element is necessary and sufficient for the induction of yeast KAR2 (BiP) gene by unfolded proteins Mori,K.A.Sant;K.Kohno;K.Normington;M.J.Gething;J.F.Sambrook
  24. Nucleic Acids Res. v.24 Saccharomyces cerevisiae IRE2/HAC1 is involved in IRE1-mediated KAR2 expression Nikawa,J.;M.Akiyoshi;S.Hirata;T.Fukuda https://doi.org/10.1093/nar/24.21.4222
  25. Cell. v.99 A role for presenilin-1 in muclear accumulation of Ire1 fragments and induction of the mammalian unfolded protein response Niwa,M.;C.Sidrauski;R.J.Kaufman;P.Walter https://doi.org/10.1016/S0092-8674(00)81667-0
  26. Nucleic Acids Res. v.22 Hacl1: A novel yeast bZIP protein binding to the CRE motif is a multicopy suppressor for cdc10 mutant of Schizosaccharomyces prombe Nojima,H.;S.H.Leem;H.Araki;A.Sakai;N.Nakashima;Y.Kanaoka;Y.Ono https://doi.org/10.1093/nar/22.24.5279
  27. Curr. Opin. Cell Biol. v.13 Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals Patil,C.;P.Walter https://doi.org/10.1016/S0955-0674(00)00219-2
  28. Nucleic Acids Res. v.27 The mammalian endoplasmic reticulum stress response element consists of an evolutionarily conserved tripartite structure and intracts with a novel stress inducible complex Roy,B.;A.S.Lee https://doi.org/10.1093/nar/27.6.1437
  29. Trends Cell Biol. v.4 The unfolded-protein-response pathway in yeast Shamu,C.E.J.S.Cox;P.Walter https://doi.org/10.1016/0962-8924(94)90011-6
  30. Genes Dev. v.12 A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells Tirasophon,W.;A.A.Welihinda;R.J.Kaufman https://doi.org/10.1101/gad.12.12.1812
  31. J. Cell Sci. v.21 IRE1and efferent signaling from the endoplasmic reticulum Urano,F.;A.Bertolotti;D.Ron
  32. EMBO J. v.17 Cloning of mammalian Ire1 reveals diversity in the ER sress responses Wang,X.Z.;H.P.Harding;Y.Zhang;E.M.Jolicoeur;M.Kuroda;D.Ron https://doi.org/10.1093/emboj/17.19.5708
  33. J. Biol. Chem. v.273 dentification of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose-regulated proteins; involvement of basic-leucine zipper transcription factors Yoshida,H.;K.Haze;H.Yanagi;T.Yura;K.Mori
  34. Cell v.107 XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor Yoshida,H.;T.Masui;Yamamoto,A.;Okada.T.;K.Mori https://doi.org/10.1016/S0092-8674(01)00611-0
  35. Mol. Cell. Biol. v.20 ATF6 activated by proteolysis directly binds in the presence of NF-Y (CBF) to the cis-acting element responsible for the mammalian unfolded protein response Yoshida,H.;T.Okada;K.Haze;H.Yanagi;T.Yura;K.Mori https://doi.org/10.1128/MCB.20.18.6755-6767.2000