Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Isolation and Characterization of a Novel Transcription Factor ATFC Activated by ER Stress from Bombyx mori Bm5 Cell Lines

누에 배양세포(Bm5)로부터 분리한 새로운 전사제어인자 ATFC의 특성분석

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

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.

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

Keywords

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