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The IRE1 pathway regulates honey bee Unfolded Protein Response gene expression.
Insect Biochemistry and Molecular Biology ( IF 3.2 ) Pub Date : 2020-03-27 , DOI: 10.1016/j.ibmb.2020.103368
Tara Reyes Adames 1 , Nicole C Rondeau 1 , Mosammed T Kabir 1 , Brittany A Johnston 2 , Henry Truong 1 , Jonathan W Snow 1
Affiliation  

Our molecular understanding of honey bee cellular stress responses is incomplete. Previously, we sought to identify and began functional characterization of the components of the Unfolded Protein Response (UPR) in honey bees. We observed that UPR stimulation resulted in induction of target genes upon IRE1 pathway activation, as assessed by splicing of Xbp1 mRNA. However, we were not able to determine the relative role of the various UPR pathways in gene activation. Our understanding of honey bee signal transduction and transcriptional regulation has been hampered by a lack of tools. After using RNA-seq to expand the known UPR targets in the honey bee, we used the Drosophila melanogaster S2 cell line and honey bee trans and cis elements to investigate the role of the IRE1 pathway in the transcriptional activation of one of these targets, the honey bee Hsc70-3 gene. Using a luciferase reporter, we show that honey bee Hsc70 promoter activity is inducible by UPR activation. In addition, we show that this activation is IRE1-dependent and relies on specific cis regulatory elements. Experiments using exogenous honey bee or fruit fly XBP1S proteins demonstrate that both factors can activate the Hsc70-3 promoter and further support a role for the IRE1 pathway in control of Hsc70-3 expression in the honey bee. By providing foundational knowledge about the UPR in the honey bee and demonstrating the usefulness of a heterologous cell line for molecular characterization of honey bee pathways, this work stands to improve our understanding of this critical species.

中文翻译:

IRE1途径调节蜜蜂未折叠蛋白反应基因的表达。

我们对蜜蜂细胞应激反应的分子理解是不完整的。以前,我们试图确定并开始对蜜蜂中未折叠蛋白反应(UPR)的成分进行功能表征。我们观察到,UPR刺激在IRE1途径激活后导致靶基因的诱导,如通过Xbp1 mRNA的剪接所评估的。但是,我们无法确定各种UPR途径在基因激活中的相对作用。缺乏工具阻碍了我们对蜜蜂信号转导和转录调控的理解。在使用RNA-seq扩展了蜜蜂中已知的UPR靶标后,我们使用了果蝇S2细胞系以及蜜蜂的反式和顺式元件来研究IRE1途径在这些靶标之一的转录激活中的作用,蜜蜂Hsc70-3基因。使用荧光素酶报道基因,我们显示了蜜蜂Hsc70启动子活性可通过UPR激活诱导。此外,我们表明这种激活是IRE1依赖的,并依赖于特定的顺式调控元件。使用外源蜜蜂或果蝇XBP1S蛋白进行的实验表明,这两个因子均可激活Hsc70-3启动子,并进一步支持IRE1途径在控制蜜蜂中Hsc70-3表达中的作用。通过提供有关蜜蜂中UPR的基础知识,并证明异源细胞系对蜜蜂途径的分子表征的有用性,这项工作旨在增进我们对这一关键物种的了解。此外,我们表明这种激活是IRE1依赖的,并依赖于特定的顺式调控元件。使用外源蜜蜂或果蝇XBP1S蛋白进行的实验表明,这两个因子均可激活Hsc70-3启动子,并进一步支持IRE1途径在控制蜜蜂中Hsc70-3表达中的作用。通过提供有关蜜蜂中UPR的基础知识,并证明异源细胞系对蜜蜂途径的分子表征的有用性,这项工作旨在增进我们对这一关键物种的了解。此外,我们表明这种激活是IRE1依赖的,并依赖于特定的顺式调控元件。使用外源蜜蜂或果蝇XBP1S蛋白进行的实验表明,这两个因子均可激活Hsc70-3启动子,并进一步支持IRE1途径在控制蜜蜂中Hsc70-3表达中的作用。通过提供有关蜜蜂中UPR的基础知识,并证明异源细胞系对蜜蜂途径的分子表征的有用性,这项工作旨在增进我们对这一关键物种的了解。使用外源蜜蜂或果蝇XBP1S蛋白进行的实验表明,这两个因子均可激活Hsc70-3启动子,并进一步支持IRE1途径在控制蜜蜂中Hsc70-3表达中的作用。通过提供有关蜜蜂中UPR的基础知识,并证明异源细胞系对蜜蜂途径的分子表征的有用性,这项工作旨在增进我们对这一关键物种的了解。使用外源蜜蜂或果蝇XBP1S蛋白进行的实验表明,这两个因子均可激活Hsc70-3启动子,并进一步支持IRE1途径在控制蜜蜂中Hsc70-3表达中的作用。通过提供有关蜜蜂中UPR的基础知识,并证明异源细胞系对蜜蜂途径的分子表征的有用性,这项工作旨在增进我们对这一关键物种的了解。
更新日期:2020-03-28
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