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3H-31, A Non-structural Protein of Heliothis virescens ascovirus 3h, Inhibits the Host Larval Cathepsin and Chitinase Activities

Virologica Sinica volume 36, pages 1036–1051 (2021)Cite this article

Abstract

3h-31 of Heliothis virescens ascovirus 3h (HvAV-3h) is a highly conserved gene of ascoviruses. As an early gene of HvAV-3h, 3h-31 codes for a non-structural protein (3H-31) of HvAV-3h. In the study, 3h-31 was initially transcribed and expressed at 3 h post-infection (hpi) in the infected Spodoptera exigua fat body cells (SeFB). 3h-31 was further inserted into the bacmid of Autographa californica nucleopolyhedrovirus (AcMNPV) to generate an infectious baculovirus (AcMNPV-31). In vivo experiments showed that budded virus production and viral DNA replication decreased with the expression of 3H-31, and lucent tubular structures were found around the virogenic stroma in the AcMNPV-31-infected SeFB cells. In vivo, both LD50 and LD90 values of AcMNPV-31 were significantly higher than those of the wild-type AcMNPV (AcMNPV-wt) in third instar S. exigua larvae. An interesting finding was that the liquefaction of the larvae killed by the infection of AcMNPV-31 was delayed. Chitinase and cathepsin activities of AcMNPV-31-infected larvae were significantly lower than those of AcMNPV-wt-infected larvae. The possible regulatory function of the chitinase and cathepsin for 3H-31 was further confirmed by RNAi, which showed that larval cathepsin activity was significantly upregulated, but chitinase activity was not significantly changed due to the RNAi of 3h-31. Based on the obtained results, we assumed that the function of 3H-31 was associated with the inhibition of host larval chitinase and cathepsin activities, so as to restrain the hosts in their larval stages.

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Acknowledgements

We thank that Dr. Qilian Qin (Institute of Zoology, Chinese Academy of Sciences) provided the SeFB cells (IOZCAS-SpexII-A), Prof. Zhihong Hu (Wuhan Institute of Virology, Chinese Academy of Sciences) provided the Escherichia coli strain DH10B and Prof. Jianhong Li (College of Plant Science & Technology, Huazhong Agricultural University) provided Ha-E cells. This research was supported by the National Natural Science Foundation of China (32070168, 31700141, 31872027) and Provincial Natural Science Foundation of Hunan (2019JJ50234), and Changsha Science and Technology Project (kq1901033), and Double first-class construction project of Hunan Agricultural University, and Sakura Science Plan of Japan Science Technology Agency (JST).

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Authors and Affiliations

  1. Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha, 410128, China

    Huan Yu, Yi-Yi Ou-Yang, Chang-Jin Yang, Ni Li & Guo-Hua Huang

  2. College of Plant Protection, Hunan Agricultural University, Changsha, 410128, China

    Huan Yu, Yi-Yi Ou-Yang, Chang-Jin Yang, Ni Li & Guo-Hua Huang

  3. Tokyo University of Agriculture and Technology, Saiwai, Fuchu, Tokyo, 183-8509, Japan

    Madoka Nakai

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  1. Huan Yu
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  2. Yi-Yi Ou-Yang
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  4. Ni Li
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Contributions

HY and GHH conceptualized and designed the study. HY, YYOY, C-JY and NL performed the experiments in the study. HY and GHH contributed reagents to the study. HY and YYOY analyzed the data. HY, MN and GHH wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Guo-Hua Huang.

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Yu, H., Ou-Yang, YY., Yang, CJ. et al. 3H-31, A Non-structural Protein of Heliothis virescens ascovirus 3h, Inhibits the Host Larval Cathepsin and Chitinase Activities. Virol. Sin. 36, 1036–1051 (2021). https://doi.org/10.1007/s12250-021-00374-y

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Keywords

  • Cathepsin
  • Chitinase
  • Heliothis virescens ascovirus 3h (HvAV-3h)
  • Recombinant virus
  • 3h-31