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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.1
- 수록면
- 783 - 793 (11page)
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초록· 키워드
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When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of ATF6α by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human ATF6α N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-hATF6α deletion variant was cleaved to liberate active transcription activator encompassing GV-hATF6α fragment which could translocate into the nucleus. The translocated GV-hATF6α fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-hATF6α(333) represents an innovative tool to investigate regulated intramembrane proteolysis of ATF6α. It can substitute active pATF6(N) binding motif-based reporter cell lines.
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참고문헌 (33)
참고문헌 신청
Back, S. H. / 2012 / Endoplasmic reticulum stress and type 2 diabetes / Annu. Rev. Biochem 81:767~793
Back, S. H. / 2006 / Cytoplasmic IRE1alpha-mediated XBP1 mRNA splicing in the absence of nuclear processing and endoplasmic reticulum stress / J. Biol. Chem 281:18691~18706
Bailey, D. / 2007 / Transmembrane bZIP transcription factors in ER stress signaling and the unfolded protein response / Antioxid. Redox Signal 9:2305~2321
Bommiasamy, H. / 2009 / ATF6alpha induces XBP1-independent expansion of the endoplasmic reticulum / J. Cell Sci 122:1626~1636
Chen, X. / 2002 / The luminal domain of ATF6senses endoplasmic reticulum(ER)stress and causes translocation of ATF6from the ER to the Golgi / J. Biol. Chem 277:13045~13052
Back, S. H. / 2006 / Cytoplasmic IRE1alpha-mediated XBP1 mRNA splicing in the absence of nuclear processing and endoplasmic reticulum stress / J. Biol. Chem 281:18691~18706
Bailey, D. / 2007 / Transmembrane bZIP transcription factors in ER stress signaling and the unfolded protein response / Antioxid. Redox Signal 9:2305~2321
Bommiasamy, H. / 2009 / ATF6alpha induces XBP1-independent expansion of the endoplasmic reticulum / J. Cell Sci 122:1626~1636
Chen, X. / 2002 / The luminal domain of ATF6senses endoplasmic reticulum(ER)stress and causes translocation of ATF6from the ER to the Golgi / J. Biol. Chem 277:13045~13052
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