DAO 125:207-215 (2017)  -  DOI: https://doi.org/10.3354/dao03144

Interaction of heat shock protein 60 (HSP60) with microRNA in Chinese mitten crab during Spiroplasma eriocheiris infection

Ming-Xiao Ning1,*, Yun-Ji Xiu1,*, Jing-Xiu Bi1, Yu-Han Liu1, Li-Bo Hou1, Zheng-Feng Ding2, Wei Gu1,3, Wen Wang1, Qing-Guo Meng1,3,**

1Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, Nanjing 210023, PR China
2Jiangsu Key Laboratory for Biofunctional Molecules, College of Life Sciences and Chemistry, Jiangsu Second Normal University, 77 West Beijing Road, Nanjing 210013, PR China
3Co-Innovation Center for Marine Bio-Industry Technology of Jiangsu Province, Lianyungang, Jiangsu 222005, PR China
*These authors contributed equally to this paper
**Corresponding author:

ABSTRACT: Heat shock protein 60 from the Chinese mitten crab Eriocheir sinensis (EsHSP60) was previously identified in relation to Spiroplasma eriocheiris infection by isobaric tags for relative and absolute quantitation labelling followed by liquid chromatography-tandem mass spectrometry. In the present study, to validate the immune function of this protein, the cDNA of the EsHSP60 gene was cloned. Various crab tissues were assessed using real-time PCR, which showed that EsHSP60 transcription occurred in all tissues examined. The expression profiles of EsHSP60 in haemolymph at transcription and protein levels when infected with S. eriocheiris were investigated by real-time PCR and Western blot analysis, respectively. A significant increase of EsHSP60 transcription and protein expression appeared post-injection in response to S. eriocheiris infection when compared to the control group. The double-luciferase reporter gene assay showed that the microRNA PC-533-3p interacted with the 3’-untranslated region of EsHSP60 and inhibited the translation of EsHSP60. The expression profiles of PC-533-3p during S. eriocheiris infection were also investigated by real-time PCR. However, the change tendency of PC-533-3p was opposite to that of the EsHSP60 after S. eriocheiris challenge. These data indicate that the EsHSP60 proteins may play an important role in mediating the immune responses of E. sinensis to an S. eriocheiris challenge.

KEY WORDS: Eriocheir sinensis · HSP60 · Spiroplasma eriocheiris stimulation

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Cite this article as: Ning MX, Xiu YJ, Bi JX, Liu YH and others (2017) Interaction of heat shock protein 60 (HSP60) with microRNA in Chinese mitten crab during Spiroplasma eriocheiris infection. Dis Aquat Org 125:207-215. https://doi.org/10.3354/dao03144

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