A Drosophila Reporter for the Translational Activation of ATF4 Marks Stressed Cells during Development.
A Drosophila Reporter for the Translational Activation of ATF4 Marks Stressed Cells during Development.
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DOI：
10.1371/journal.pone.0126795
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kang MJ
中科院分区：
文献类型：
--
作者：
Kang K;Ryoo HD;Park JE;Yoon JH;Kang MJ
真核细胞已经进化出有助于恢复细胞稳态以响应各种生理或病理条件的信号通路。ATF4是一种转录因子，其mRNA翻译响应于应激激活的eIF2 α激酶而被刺激。激活eIF2 α磷酸化和ATF4翻译的既定条件包括内质网（ER）中的过度应激和氨基酸剥夺。ATF 4通过一种独特的翻译激活机制被激活，该机制涉及5 '非翻译区（UTR）中的多个上游开放阅读框（uORF），该非翻译区从酵母到哺乳动物都是保守的。利用这一点，我们开发了果蝇中ATF 4的翻译激活报告基因，其中dsRed报告基因编码序列位于果蝇ATF 4 5' UTR的下游。在正常条件下，该报告基因在大多数组织中保持无活性，但在饥饿时或在施加ER应激的条件下激发时显示dsRed表达。在正常发育的果蝇中，即使没有外源性应激，少数细胞类型也显示报告基因表达，包括唾液腺、肠道、雄性生殖器官和感光细胞，这表明这些细胞类型在正常发育期间存在内在应激。这些结果为研究ATF4介导的果蝇发育和疾病中的应激反应提供了新的工具。
Eukaryotic cells have evolved signaling pathways that help to restore cellular homeostasis in response to various physiological or pathological conditions. ATF4 is a transcription factor whose mRNA translation is stimulated in response to stress-activated eIF2alpha kinases. Established conditions that activate eIF2alpha phosphorylation and ATF4 translation include excessive stress in the endoplasmic reticulum (ER) and amino acid deprivation. ATF4 is activated through a unique translational activation mechanism that involves multiple upstream open reading frames (uORFs) in the 5’-untranslated region (UTR), which is conserved from yeast to mammals. Taking advantage of this, we developed a translational activation reporter of ATF4 in Drosophila, in which the dsRed reporter coding sequence was placed downstream of the Drosophila ATF4 5’ UTR. This reporter remained inactive in most tissues under normal conditions, but showed dsRed expression when starved, or when challenged with conditions that imposed ER stress. In normally developing flies, a small number of cell types showed reporter expression even without exogenous stress, which included the salivary gland, gut, the male reproductive organ, and the photoreceptor cells, suggestive of inherent stress during the normal development of these cell types. These results establish a new tool to study ATF4-mediated stress response in Drosophila development and disease.
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