Disease-related amyloidogenic variants of human lysozyme trigger the unfolded protein response and disturb eye development in Drosophila melanogaster.
Disease-related amyloidogenic variants of human lysozyme trigger the unfolded protein response and disturb eye development in Drosophila melanogaster.
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DOI：
10.1096/fj.11-185983
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发表时间：
2012-01
期刊：
影响因子：
--
通讯作者：
Brorsson AC
中科院分区：
文献类型：
--
作者：
Kumita JR;Helmfors L;Williams J;Luheshi LM;Menzer L;Dumoulin M;Lomas DA;Crowther DC;Dobson CM;Brorsson AC
我们已经创建了一个果蝇模型的溶菌酶淀粉样变性研究疾病相关变异的体内行为。为了实现这一目标，野生型（WT）蛋白和淀粉样变异体F57I和D67H在黑腹果蝇中使用UAS-gal4系统以及普遍存在和视网膜表达驱动因子Act5C-gal4和gmr-gal4表达。非转基因w1118果蝇系作为对照。我们利用ELISA实验检测溶菌酶蛋白水平，扫描电镜检测眼表型分类，免疫组织化学检测未折叠蛋白反应（UPR）激活。我们观察到，表达不稳定的F57I和D67H溶菌酶会触发UPR激活，导致这些变体的降解，而WT溶菌酶则分泌到苍蝇的血淋巴中。事实上，WT的含量是变异蛋白的17倍。此外，F57I变异引起眼睛发育的显著中断，这与明显的UPR激活相关。这些结果支持这样一个概念，即家族性淀粉样蛋白疾病的发病与UPR在分泌前无法完全降解淀粉样蛋白溶菌酶有关，从而导致这些不稳定变体的分泌，从而导致沉积和相关的器官损伤。-Kumita, j.r, Helmfors， L., Williams， J., Luheshi， L. M., Menzer， L., Dumoulin， M., Lomas， D., Crowther， D., Dobson， C. M., brosson， A.-疾病相关的人类溶菌酶淀粉样变异体触发未折叠蛋白反应并干扰黑腹果蝇的眼睛发育。
We have created a Drosophila model of lysozyme amyloidosis to investigate the in vivo behavior of disease-associated variants. To achieve this objective, wild-type (WT) protein and the amyloidogenic variants F57I and D67H were expressed in Drosophila melanogaster using the UAS-gal4 system and both the ubiquitous and retinal expression drivers Act5C-gal4 and gmr-gal4. The nontransgenic w1118 Drosophila line was used as a control throughout. We utilized ELISA experiments to probe lysozyme protein levels, scanning electron microscopy for eye phenotype classification, and immunohistochemistry to detect the unfolded protein response (UPR) activation. We observed that expressing the destabilized F57I and D67H lysozymes triggers UPR activation, resulting in degradation of these variants, whereas the WT lysozyme is secreted into the fly hemolymph. Indeed, the level of WT was up to 17 times more abundant than the variant proteins. In addition, the F57I variant gave rise to a significant disruption of the eye development, and this correlated to pronounced UPR activation. These results support the concept that the onset of familial amyloid disease is linked to an inability of the UPR to degrade completely the amyloidogenic lysozymes prior to secretion, resulting in secretion of these destabilized variants, thereby leading to deposition and associated organ damage.—Kumita, J. R., Helmfors, L., Williams, J., Luheshi, L. M., Menzer, L., Dumoulin, M., Lomas, D. A., Crowther, D. C., Dobson, C. M., Brorsson, A.-C. Disease-related amyloidogenic variants of human lysozyme trigger the unfolded protein response and disturb eye development in Drosophila melanogaster.
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影响因子：
2.9
作者：
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通讯作者：
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