Zhang Congfen, Dai Lishang, Wang Lei, Qian Cen, Wei Guoqing, Li Jun, Zhu Baojian, Liu Chaoliang
College of Life Science, Anhui Agricultural University, Anhui Hefei, P.R. China, 230036; Department of Pharmacology, Wannan Medical College, Anhui Wuhu, P.R.China, 241002.
College of Life Science, Anhui Agricultural University, Anhui Hefei, P.R. China, 230036.
PLoS One. 2015 Apr 6;10(4):e0121296. doi: 10.1371/journal.pone.0121296. eCollection 2015.
Small heat shock proteins (sHSPs) can regulate protein folding and protect cells from stress. To investigate the role of sHSPs in the silk-producing insect Antheraea pernyi response to microorganisms, a sHsp gene termed as Ap-sHSP21.4, was identified. This gene encoded a 21.4 kDa protein which shares the conserved structure of insect sHsps and belongs to sHSP21.4 family. Ap-sHSP21.4 was highly expressed in fat body and up-regulated in midgut and fat body of A. pernyi challenged with Escherichia coli, Beauveria bassiana and nuclear polyhedrosis virus (NPV), which was determined by quantitative real-time PCR. Meanwhile, knock down of Ap-sHSP21.4 with dsRNA result in the decrease at the expression levels of several immune response-related genes (defensin, Dopa decarboxylase, Toll1, lysozyme and Kazal-type serine protease inhibitor). Additionally, the impact of eicosanoid biosynthesis on the expression of Ap-sHSP21.4 response to NPV was determined using qPCR, inhibitors of eicosanoid biosynthesis significantly suppress Ap-HSP21.4 expression upon NPV challenge. All together, Ap-sHSP21.4 was involved in the immunity of A. pernyi against microorganism and possibly mediated by eicosanoids pathway. These results will shed light in the understanding of the pathogen-host interaction in A. pernyi.
小热休克蛋白(sHSPs)可以调节蛋白质折叠并保护细胞免受应激。为了研究sHSPs在产丝昆虫柞蚕对微生物反应中的作用,鉴定了一个名为Ap-sHSP21.4的sHsp基因。该基因编码一种21.4 kDa的蛋白质,其具有昆虫sHsps的保守结构,属于sHSP21.4家族。通过定量实时PCR测定,Ap-sHSP21.4在脂肪体中高度表达,并且在受到大肠杆菌、球孢白僵菌和核型多角体病毒(NPV)攻击的柞蚕中肠和脂肪体中上调。同时,用dsRNA敲低Ap-sHSP21.4导致几种免疫反应相关基因(防御素、多巴脱羧酶、Toll1、溶菌酶和Kazal型丝氨酸蛋白酶抑制剂)的表达水平下降。此外,使用qPCR测定了类二十烷酸生物合成对Ap-sHSP21.4对NPV反应表达的影响,类二十烷酸生物合成抑制剂在NPV攻击后显著抑制Ap-HSP21.4的表达。总之,Ap-sHSP21.4参与了柞蚕对微生物的免疫,并且可能由类二十烷酸途径介导。这些结果将有助于理解柞蚕中病原体与宿主的相互作用。