Analysis of the UPR, a transcriptional induction process coupled with intracellular signaling from the endoplasmic reticulum to the nucleus

UPR 分析，这是一个与从内质网到细胞核的细胞内信号传导相结合的转录诱导过程

• 批准号: 12680692
• 负责人: MORI Kazutoshi
• 金额: $ 2.37万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680692/
• 关键词: endoplasmic reticulum; molecular chaperone; folding enzymes; intracellular signaling; transcriptional induction; transcription factor; proteolysis; nuclear translocation; 転写因子; マイクロアレー; アルツハイマー病; プレセニリン; フォールディング

By 1999, we showed that the human basic leucine zipper protein ATF6 we isolated as a transcription factor specific to the mammalian UPR is constitutively synthesized as a type II transmembrane glycoprotein localized in the endoplasmic reticulum (ER) and activated by proteolysis in response to ER stress. From 2000 to 2001, we further analyzed various properties of ATF6 in more detail and obtained the following results.(1) The N-terminal fragment of ATF6 relocates from the ER to the nucleus upon ER stress-induced proteolysis (visualized by indirect immunofluorescence analysis). (2) The consensus sequence of the cis-acting ER stress response element (ERSE) necessary and sufficient for the mammalian UPR is CCAAT-N9-CCACG. Although ATF6 cannot bind to ERSE by itself, ATF6 can bind to the CCACG part of ERSE when the CCAAT part is bound to the ubiquitous transcription factor NF-Y. (3) Transcriptional activities of wild-type and various mutant ERSE-like sequences are correlated well with their abilities to bind ATF6. (4) ER stress-induced proteolysis of ATF6 and subsequent transcriptional induction of BiP/GRP78 (a major molecular chaperone in the ER) is specifically blocked by the serine protease inhibitor AEBSF at the concentration of 300 μM. (5) Comprehensive analysis of target genes of the UPR and ATF6 revealed that approximately half of UPR-target genes are directly regulated by ATF6 and that a majority of ATF6-target genes encodes molecular chaperones and folding enzymes in the ER. (6) In cells possessing Familial Alzheimer disease-linked mutation in the presenilin gene, ER stress-induced activation of ATF6 is mitigated.Above results strongly indicate that ATF6 plays a major role in the mammalian UPR.

到1999年，我们发现，作为哺乳动物UPR特异性转录因子，我们分离的人碱性亮氨酸拉链蛋白ATF6组成性合成为II型跨膜糖蛋白，定位于内质网（ER），并在内质网应激下被蛋白水解激活。从2000年到2001年，我们进一步详细分析了ATF6的各种性能，得到了以下结果：(1)在内质网应激诱导的蛋白水解作用下，ATF6的n端片段从内质网迁移到细胞核（通过间接免疫荧光分析可见）。(2)哺乳动物UPR必需和充分的顺式内质网应激反应元件（ERSE）的共识序列是CCAAT-N9-CCACG。虽然ATF6本身不能与ERSE结合，但当CCAAT部分与无处不在的转录因子NF-Y结合时，ATF6可以与ERSE的CCACG部分结合。(3)野生型和各种突变型erse样序列的转录活性与其结合ATF6的能力密切相关。(4) 300 μM的丝氨酸蛋白酶抑制剂AEBSF特异性阻断内质网应激诱导的ATF6蛋白水解和随后的BiP/GRP78（内质网中的主要分子伴侣）的转录诱导。(5)综合分析UPR和ATF6的靶基因发现，大约一半的UPR靶基因直接受ATF6调控，ATF6靶基因大部分编码内质网中的分子伴侣蛋白和折叠酶。(6)在具有家族性阿尔茨海默病相关早老素基因突变的细胞中，内质网应激诱导的ATF6激活被减轻。以上结果有力地表明，ATF6在哺乳动物UPR中起着重要作用。

项目成果

Kyosuke Haze: "Identification of the G13 (cAMP response element binding protein-related protein) gene product related to ATF6 as a transcriptional activator"Biochemical Journal. 354(in press). (2001)

Kyosuke Haze：“作为转录激活剂与 ATF6 相关的 G13（cAMP 反应元件结合蛋白相关蛋白）基因产物的鉴定”生化杂志。

Katayama, Imaizumi, Honda, Yoneda, Kudo, Takeda, Mori, Rozmahel, Franser, George-Hyslop, Tohyama: "Disturbed Activation of Endoplasmic Reticulum Stress Transducers by Familial Alzheimer's Disease-linked Presenilin-1 Mutations"THE JOURNAL OF BIOLOGICAL CHE

Katayama、Imaizumi、Honda、Yoneda、Kudo、Takeda、Mori、Rozmahel、Franser、George-Hyslop、Tohyama：“家族性阿尔茨海默病相关的 Presenilin-1 突变对内质网应激传感器的激活干扰”生物化学杂志

Hiderou Yoshida: "Endoplasmic reticulum stress-induced formation of transcription factor complex ERSF including NF-Y(CBF) and activationg transcription factors 6α and 6β"Molecular and Cellular Biology. Vol.21 No.4. 1239-1248 (2001)

Hiderou Yoshida：“内质网应激诱导转录因子复合物ERSF的形成，包括NF-Y(CBF)和激活转录因子6α和6β”《分子和细胞生物学》第21卷第4期(2001)。

Hiderou Yoshida: "ATF6 activated by proteolysis binds in the presence of NF-Y(CBF) directly to the cis-acting element responsible for the mammalian unfolded protein response"Molecular and Cellular Biology. 20. 6755-6767 (2000)

Hiderou Yoshida：“在 NF-Y(CBF) 存在的情况下，通过蛋白水解激活的 ATF6 直接与负责哺乳动物未折叠蛋白反应的顺式作用元件结合”《分子和细胞生物学》。

Taiichi Katayama: "Disturbed Activation of Endoplasmic Reticulum Stress Transducers by Familial Alzheimer's Disease-linked Presenilin-1 Mutations"Journal of Blological Chemistry. 276. 43446-43454 (2001)

Taiichi Katayama：“家族性阿尔茨海默氏病相关的 Presenilin-1 突变对内质网应激传感器的激活干扰”，《生物化学杂志》。