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鉴定和功能的 cAMP 反应元件结合蛋白在 Oak 蚕天蚕。

Identification and function of cAMP response element binding protein in Oak silkworm Antheraea pernyi.

机构信息

College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.

College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.

出版信息

J Invertebr Pathol. 2018 Jan;151:14-20. doi: 10.1016/j.jip.2017.10.006. Epub 2017 Oct 24.

Abstract

Cyclic AMP response element binding (CREB) proteins participate in the regulation of many biological processes. However, little is known about their role in immune regulation in the Oak silkworm (Antheraea pernyi). In this study, a CREB gene was identified in A. pernyi and its role in immune regulation was investigated. ApCREB shares conserved signature motifs with other CREB proteins, and includes a typical leucine zipper domain, specific DNA-binding site, nuclear localisation signal (NLS) and cAMP-dependent protein kinase phosphorylation site. Recombinant ApCREB was expressed in Escherichia coli and used to raise rabbit anti-ApCREB polyclonal antibodies. ApCREB mRNA was detected in all examined tissues, with maximum expression in the midgut and integument. Following exposure to four pathogenic microorganisms (Beauveria bassiana, Escherichia coli, Micrococcus luteus, and Antheraea pernyi nuclear polyhedrosis virus), expression of ApCREB was up-regulated by B. bassiana, E. coli and ApNPV, down-regulated by M. luteus. RNA interference of ApCREB affected mRNA expression levels of antimicrobial peptide genes attacin-1, cecropin B, defensin-1, gloverin, and lebocin-2. These findings demonstrate that ApCREB is a CREB homologue that may be involved in innate immunity in A. pernyi.

摘要

环磷酸腺苷反应元件结合蛋白(CREB)参与许多生物过程的调节。然而,它们在柞蚕(Antheraea pernyi)免疫调节中的作用知之甚少。本研究在柞蚕中鉴定了一个 CREB 基因,并研究了其在免疫调节中的作用。ApCREB 与其他 CREB 蛋白具有保守的特征基序,包括典型的亮氨酸拉链结构域、特定的 DNA 结合位点、核定位信号(NLS)和 cAMP 依赖性蛋白激酶磷酸化位点。重组 ApCREB 在大肠杆菌中表达,并用于制备兔抗 ApCREB 多克隆抗体。在所有检测的组织中均检测到 ApCREB mRNA 的表达,其中在中肠和表皮中表达量最高。在暴露于四种致病性微生物(球孢白僵菌、大肠杆菌、藤黄微球菌和柞蚕核型多角体病毒)后,ApCREB 的表达被球孢白僵菌、大肠杆菌和 ApNPV 上调,被藤黄微球菌下调。ApCREB 的 RNA 干扰影响了抗菌肽基因 attacin-1、cecropin B、defensin-1、gloverin 和 lebocin-2 的 mRNA 表达水平。这些发现表明,ApCREB 是一种 CREB 同源物,可能参与柞蚕的先天免疫。

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