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果蝇未折叠蛋白反应 (UPR) 的体外和体内检测。

Drosophila Unfolded Protein Response (UPR) Assays In Vitro and In Vivo.

机构信息

Department of Cell Biology, New York University Grossman School of Medicine, New York, NY, USA.

出版信息

Methods Mol Biol. 2022;2378:261-277. doi: 10.1007/978-1-0716-1732-8_17.

Abstract

Wildtype or mutant proteins expressed beyond the capacity of a cell's protein folding system could be detrimental to general cellular function and survival. In response to misfolded protein overload in the endoplasmic reticulum (ER), eukaryotic cells activate the Unfolded Protein Response (UPR) that helps cells restore protein homeostasis in the endoplasmic reticulum (ER). As part of the UPR, cells attenuate general mRNA translation and activate transcription factors that induce stress-responsive gene expression.UPR signaling draws research interest in part because conditions that cause chronic protein misfolding in the ER or those that impair UPR signaling underlie several diseases including neurodegeneration, diabetes, and cancers. Model organisms are frequently employed in the field as the UPR pathways are generally well-conserved throughout phyla. Here, we introduce experimental procedures to detect UPR in Drosophila melanogaster.

摘要

野生型或突变蛋白的表达超过细胞蛋白质折叠系统的能力可能对一般细胞功能和生存有害。为了应对内质网(ER)中错误折叠蛋白的过载,真核细胞激活未折叠蛋白反应(UPR),帮助细胞在内质网(ER)中恢复蛋白质的平衡。作为 UPR 的一部分,细胞减弱一般的 mRNA 翻译,并激活转录因子,诱导应激反应基因的表达。UPR 信号引起了研究兴趣,部分原因是导致 ER 中慢性蛋白质错误折叠的条件,或那些破坏 UPR 信号的条件,是几种疾病的基础,包括神经退行性疾病、糖尿病和癌症。模式生物在该领域经常被使用,因为 UPR 途径在整个门中通常是保守的。在这里,我们介绍了检测黑腹果蝇中 UPR 的实验程序。

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