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两种丝裂原活化蛋白激酶的分子特征:家蚕(鳞翅目:蚕蛾科)的p38丝裂原活化蛋白激酶和核糖体S6激酶,以及对它们在应对家蚕核型多角体病毒感染中作用的深入了解。

Molecular Characterization of Two Mitogen-Activated Protein Kinases: p38 MAP Kinase and Ribosomal S6 Kinase From Bombyx mori (Lepidoptera: Bombycidae), and Insight Into Their Roles in Response to BmNPV Infection.

作者信息

Muhammad Azharuddin, Toufeeq Shahzad, Yu Hai-Zhong, Wang Jie, Zhang Shang-Zhi, Li Bing, Li Zhen, Yang Li-Ang, Hu Pei, Ma Yan, Xu Jia-Ping

机构信息

School of Life Sciences, Anhui Agricultural University, Hefei, China.

Anhui International Joint Research and Developmental Center of Sericulture Resources Utilization, Hefei, China.

出版信息

J Insect Sci. 2019 Jan 1;19(1):15. doi: 10.1093/jisesa/iey134.

DOI:10.1093/jisesa/iey134
PMID:30715437
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6359879/
Abstract

Proteins p38 map kinase and ribosomal S6 kinase (S6K) as members of mitogen-activated protein kinases (MAPKs) play important roles against pathogens. In this study, Bmp38 and BmS6K were identified as differentially expressed proteins from iTRAQ database. Bmp38 and BmS6K were expressed, and recombinant proteins were purified. The bioinformatics analysis showed that both proteins have serine/threonine-protein kinases, catalytic domain (S_TKc) with 360 and 753 amino acids, respectively. The real-time quantitative polymerase chain reaction (RT-qPCR) results suggest that Bmp38 and BmS6K had high expression in the midgut and hemolymph. The comparative expression level of Bmp38 and BmS6K in BC9 was upregulated than in P50 in the midgut after Bombyx mori nucleopolyhedrovirus (BmNPV) infection. Western bolt results showed a positive correlation between RT-qPCR and iTRAQ data for Bmp38, but BmS6K data showed partial correlation with iTRAQ. Injection of anti-Bmp38 and anti-BmS6K serum suggested that Bmp38 may be involved against BmNPV infection, whereas BmS6K may require phosphorylation modification to inhibit BmNPV infection. Taken together, our results suggest that Bmp38 and BmS6k might play an important role in innate immunity of silkworm against BmNPV.

摘要

蛋白质p38丝裂原活化蛋白激酶和核糖体S6激酶(S6K)作为丝裂原活化蛋白激酶(MAPK)家族的成员,在抵抗病原体方面发挥着重要作用。在本研究中,Bmp38和BmS6K被鉴定为来自iTRAQ数据库的差异表达蛋白。对Bmp38和BmS6K进行了表达,并纯化了重组蛋白。生物信息学分析表明,这两种蛋白均具有丝氨酸/苏氨酸蛋白激酶催化结构域(S_TKc),分别含有360和753个氨基酸。实时定量聚合酶链反应(RT-qPCR)结果表明,Bmp38和BmS6K在中肠和血淋巴中高表达。家蚕核型多角体病毒(BmNPV)感染后,BC9中肠中Bmp38和BmS6K的相对表达水平高于P50。蛋白质免疫印迹结果显示,Bmp38的RT-qPCR数据与iTRAQ数据呈正相关,但BmS6K的数据与iTRAQ呈部分相关。注射抗Bmp38和抗BmS6K血清表明,Bmp38可能参与抵抗BmNPV感染,而BmS6K可能需要磷酸化修饰才能抑制BmNPV感染。综上所述,我们的结果表明,Bmp38和BmS6k可能在蚕对BmNPV的先天免疫中发挥重要作用。

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