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Characterization of lamin B receptor of Sf9 cells and its fate during Autographa californica nucleopolyhedrovirus infection

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Abstract

Baculovirus nucleocapsids egress from the nuclear membrane during infection. However, details of alternation of nuclear membrane structure during baculovirus egress are unknown. In this study, we examined the changes of lamin B receptor (LBR), a main inner nuclear membrane component, during Autographa californica nucleopolyhedrovirus (AcMNPV) infection. Firstly, the open reading frame (Orf) of Sf9 lbr was cloned by reverse transcription PCR, and the distribution of LBR in Sf9 cells were observed by fusing LBR with the red fluorescence protein mcherry. Besides, the amount of endogenous LBR during AcMNPV infection was detected by western blotting. Moreover, the distribution of LBR after AcMNPV infection was observed under the confocal fluorescence microscopy. Furthermore, the effects of protein kinase C (PKC) inhibitor on stability of LBR and release of budded virus (BVs) were determined. The results showed that Sf9 lbr contains an Orf of 2040 nucleotides (NTs), which encodes a predicted protein of 679 amino acids (AAs). Fluorescence microscopy showed that LBR is localized to the nuclear membrane. Western blotting result showed that the amount of endogenous LBR is significantly reduced after AcMNPV infection. Transfection and infection assay demonstrated that the fluorescence of LBR nearly completely disappeared after viral infection. PKC inhibitor can suppress the degradation of LBR induced by AcMNPV, resulting in the reduction of viral titer of progeny viruses. The electron microscopy analysis demonstrated that PKC inhibitor did not influence virion entry, uncoating, and assembly, but may partially protect the nuclear membrane from disruption by AcMNPV. Taken together, AcMNPV infection can distort the expression of LBR, which may promote the egress of nucleocapsids.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (31501701), and the Plant Foundation for Young Scientists of Henan University (CX0000A40557). We thank all the members of our laboratory for excellent experimental assistance. We also acknowledge Aiyu Guo for his help with the confocal fluorescence microscopy.

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  1. Wenqiang Wei and Zichao Hu contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cell Signal Transduction and Institute of Biomedical Informatics, Henan University, Kaifeng, 475004, Henan, China

    Wenqiang Wei, Zichao Hu, Yuting Jia, TingXuan Gu, Wei Zhao & Shaoping Ji

  2. Henan International Joint Laboratory for Nuclear Protein in Gene Regulation, Henan University, Kaifeng, 475004, Henan, China

    Wenqiang Wei

  3. Department of Microbiology, School of Basic Medical Sciences, Henan University, Kaifeng, 475004, Henan, China

    Wenqiang Wei

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  1. Wenqiang Wei
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  2. Zichao Hu
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  5. Wei Zhao
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Correspondence to Wenqiang Wei or Shaoping Ji.

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Wei, W., Hu, Z., Jia, Y. et al. Characterization of lamin B receptor of Sf9 cells and its fate during Autographa californica nucleopolyhedrovirus infection. Cytotechnology 72, 315–325 (2020). https://doi.org/10.1007/s10616-020-00380-0

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  • DOI: https://doi.org/10.1007/s10616-020-00380-0

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