Hostname: page-component-cd9895bd7-gxg78 Total loading time: 0 Render date: 2024-12-29T05:14:59.409Z Has data issue: false hasContentIssue false
  • English
  • Français

Expression of anti-NbHK single-chain antibody in fusion with NSlmb enhances the resistance to Nosema bombycis in Sf9-III cells

Published online by Cambridge University Press:  06 April 2022

Renze Zhang ,
Shiyi Zheng ,
Hongyun Huang ,
Xi Sun ,
Yukang Huang ,
Junhong Wei ,
Guoqing Pan ,
Chunfeng Li  and
Zeyang Zhou
Renze Zhang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Shiyi Zheng
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Affiliated Jinhua Hospital, Zhejiang University of Medicine, Jinhua Municipal Central Hospital, Jinhua, Zhejiang 321000, China
Hongyun Huang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Xi Sun
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Yukang Huang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Junhong Wei
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Guoqing Pan
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Chunfeng Li*
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Zeyang Zhou
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China College of Life Sciences, Chongqing Normal University, Chongqing 401331, China
*
Author for correspondence: Chunfeng Li, Email: licf@swu.edu.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

Nosema bombycis is a destructive and specific intracellular parasite of silkworm, which is extremely harmful to the silkworm industry. N. bombycis is considered as a quarantine pathogen of sericulture because of its long incubation period and horizontal and vertical transmission. Herein, two single-chain antibodies targeting N. bombycis hexokinase (NbHK) were cloned and expressed in fusion with the N-terminal of Slmb (a Drosophila melanogaster FBP), which contains the F-box domain. Western blotting demonstrated that Sf9-III cells expressed NSlmb–scFv-7A and NSlmb–scFv-6H, which recognized native NbHK. Subsequently, the NbHK was degraded by host ubiquitination system. When challenged with N. bombycis, the transfected Sf9-III cells exhibited better resistance relative to the controls, demonstrating that NbHK is a prospective target for parasite controls and this approach represents a potential solution for constructing N. bombycis-resistant Bombyx mori.

Keywords

HexokinaseNosema bombycisresistantsingle-chain antibodyubiquitination system
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 112 , Issue 4 , August 2022 , pp. 502 - 508
Check for updates
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

*

These authors contributed equally to the work reported in this paper.

References

Caussinus, E, Kanca, O and Affolter, M (2011) Fluorescent fusion protein knockout mediated by anti-GFP nanobody. Nature Structural & Molecular Biology 19, 117–121.CrossRefGoogle ScholarPubMed
Chen, J, Geng, L, Long, M, Li, T, Li, Z, Yang, D, Ma, C, Wu, H, Ma, Z, Li, C, Pan, G and Zhou, Z (2013) Identification of a novel chitin-binding spore wall protein (NbSWP12) with a BAR-2 domain from Nosema bombycis (microsporidia). Parasitology 140, 1394–1402.CrossRefGoogle ScholarPubMed
Ciechanover, A (2005) N-terminal ubiquitination. Methods in Molecular Biology 301, 255–270.Google ScholarPubMed
Ciechanover, A and Ben-Saadon, R (2004) N-terminal ubiquitination: more protein substrates join in. Trends in Cell Biology 14, 103–106.CrossRefGoogle Scholar
Cuomo, CA, Desjardins, CA, Bakowski, MA, Goldberg, J, Ma, AT, Becnel, JJ, Didier, ES, Fan, L, Heiman, DI, Levin, JZ, Young, S, Zeng, Q and Troemel, ER (2012) Microsporidian genome analysis reveals evolutionary strategies for obligate intracellular growth. Genome Research 22, 2478–2488.CrossRefGoogle ScholarPubMed
Dean, P, Hirt, RP and Embley, TM (2016) Microsporidia: why make nucleotides if you can steal them? PLoS Pathogens 12, e1005870.CrossRefGoogle Scholar
Dui, W, Lu, W, Ma, J and Jiao, R (2012) A systematic phenotypic screen of F-box genes through a tissue-specific RNAi-based approach in Drosophila. Journal of Genetics and Genomics 39, 397–413.CrossRefGoogle ScholarPubMed
Embley, TM and Martin, W (2006) Eukaryotic evolution, changes and challenges. Nature 440, 623–630.CrossRefGoogle ScholarPubMed
Fang, W, Vega-Rodriguez, J, Ghosh, AK, Jacobs-Lorena, M, Kang, A and St Leger, RJ (2011) Development of transgenic fungi that kill human malaria parasites in mosquitoes. Science (New York, N.Y.) 331, 1074–1077.CrossRefGoogle ScholarPubMed
Ferguson, S and Lucocq, J (2019) The invasive cell coat at the microsporidian Trachipleistophora hominis-host cell interface contains secreted hexokinases. Microbiologyopen 8, e00696.CrossRefGoogle ScholarPubMed
Grumati, P and Dikic, I (2018) Ubiquitin signaling and autophagy. Journal of Biological Chemistry 293, 5404–5413.CrossRefGoogle ScholarPubMed
Hacker, C, Howell, M, Bhella, D and Lucocq, J (2014) Strategies for maximizing ATP supply in the microsporidian Encephalitozoon cuniculi: direct binding of mitochondria to the parasitophorous vacuole and clustering of the mitochondrial porin VDAC. Cellular Microbiology 16, 565–579.CrossRefGoogle Scholar
Han, B and Weiss, LM (2017) Microsporidia: obligate intracellular pathogens within the fungal kingdom. Microbiology Spectrum 5, 10.1128.CrossRefGoogle ScholarPubMed
Han, B, Ma, Y, Tu, V, Tomita, T, Mayoral, J, Williams, T, Horta, A, Huang, H and Weiss, LM (2019) Microsporidia interact with host cell mitochondria via voltage-dependent anion channels using sporoplasm surface protein 1. mBio 10, e01944-19.CrossRefGoogle ScholarPubMed
He, Q, Luo, J, Xu, JZ, Wang, CX, Meng, XZ, Pan, GQ, Li, T and Zhou, ZY (2020) Morphology and transcriptome analysis of Nosema bombycis sporoplasm and insights into the initial infection of microsporidia. mSphere 5, e00958-19.CrossRefGoogle ScholarPubMed
Heinz, E, Hacker, C, Dean, P, Mifsud, J, Goldberg, AV, Williams, TA, Nakjang, S, Gregory, A, Hirt, RP, Lucocq, JM, Kunji, ER and Embley, TM (2014) Plasma membrane-located purine nucleotide transport proteins are key components for host exploitation by microsporidian intracellular parasites. PLoS Pathogens 10, e1004547.CrossRefGoogle ScholarPubMed
Huang, Y, Chen, J, Sun, B, Zheng, R, Li, B, Li, Z, Tan, Y, Wei, J, Pan, G, Li, C and Zhou, Z (2018 a) Engineered resistance to Nosema bombycis by in vitro expression of a single-chain antibody in Sf9-III cells. PLoS One 13, e0193065.CrossRefGoogle ScholarPubMed
Huang, Y, Zheng, S, Mei, X, Yu, B, Sun, B, Li, B, Wei, J, Chen, J, Li, T, Pan, G, Zhou, Z and Li, C (2018 b) A secretory hexokinase plays an active role in the proliferation of Nosema bombycis. PeerJ 6, e5658.CrossRefGoogle ScholarPubMed
Isaacs, AT, Li, F, Jasinskiene, N, Chen, X, Nirmala, X, Marinotti, O, Vinetz, JM and James, AA (2011) Engineered resistance to Plasmodium falciparum development in transgenic Anopheles stephensi. PLoS Pathogens 7, e1002017.CrossRefGoogle ScholarPubMed
Isaacs, AT, Jasinskiene, N, Tretiakov, M, Thiery, I, Zettor, A, Bourgouin, C and James, AA (2012) Transgenic Anopheles stephensi coexpressing single-chain antibodies resist Plasmodium falciparum development. Proceedings of the National Academy of Sciences of the United States of America 109, E1922–E1930.Google ScholarPubMed
Ishihara, R (1969) The life cycle of Nosema bombycis as revealed in tissue culture cells of Bombyx mori. Journal of Invertebrate Pathology 14, 316–320.CrossRefGoogle ScholarPubMed
Katinka, MD, Duprat, S, Cornillot, E, Metenier, G, Thomarat, F, Prensier, G, Barbe, V, Peyretaillade, E, Brottier, P, Wincker, P, Delbac, F, El Alaoui, H, Peyret, P, Saurin, W, Gouy, M, Weissenbach, J and Vivares, CP (2001) Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi. Nature 414, 450–453.CrossRefGoogle ScholarPubMed
Li, Z, Pan, G, Li, T, Huang, W, Chen, J, Geng, L, Yang, D, Wang, L and Zhou, Z (2012) SWP5, a spore wall protein, interacts with polar tube proteins in the parasitic microsporidian Nosema bombycis. Eukaryotic Cell 11, 229–237.CrossRefGoogle ScholarPubMed
Nakjang, S, Williams, TA, Heinz, E, Watson, AK, Foster, PG, Sendra, KM, Heaps, SE, Hirt, RP and Martin Embley, T (2013) Reduction and expansion in microsporidian genome evolution: new insights from comparative genomics. Genome Biology and Evolution 5, 2285–2303.CrossRefGoogle ScholarPubMed
Nandi, D, Tahiliani, P, Kumar, A and Chandu, D (2006) The ubiquitin-proteasome system. Journal of Biosciences 31, 137–155.CrossRefGoogle ScholarPubMed
Pan, G, Xu, J, Li, T, Xia, Q, Liu, SL, Zhang, G, Li, S, Li, C, Liu, H, Yang, L, Liu, T, Zhang, X, Wu, Z, Fan, W, Dang, X, Xiang, H, Tao, M, Li, Y, Hu, J, Li, Z, Lin, L, Luo, J, Geng, L, Wang, L, Long, M, Wan, Y, He, N, Zhang, Z, Lu, C, Keeling, PJ, Wang, J, Xiang, Z and Zhou, Z (2013) Comparative genomics of parasitic silkworm microsporidia reveal an association between genome expansion and host adaptation. BMC Genomics 14, 186.CrossRefGoogle ScholarPubMed
Pan, G, Bao, J, Ma, Z, Song, Y, Han, B, Ran, M, Li, C and Zhou, Z (2018) Invertebrate host responses to microsporidia infections. Developmental & Comparative Immunology 83, 104–113.CrossRefGoogle ScholarPubMed
Shiflett, AM and Johnson, PJ (2010) Mitochondrion-related organelles in eukaryotic protists. Annual Review of Microbiology 64, 409–429.CrossRefGoogle ScholarPubMed
Smyk, B, Cienciala, M and Kosiek, T (1952) Diagnosis of Nosema bombycis infection. Medycyna Doswiadczalna i Mikrobiologia 4, 363–366.Google ScholarPubMed
Sumitani, M, Kasashima, K, Yamamoto, DS, Yagi, K, Yuda, M, Matsuoka, H and Yoshida, S (2013) Reduction of malaria transmission by transgenic mosquitoes expressing an antisporozoite antibody in their salivary glands. Insect Molecular Biology 22, 41–51.CrossRefGoogle ScholarPubMed
Swatek, KN and Komander, D (2016) Ubiquitin modifications. Cell Research 26, 399–422.CrossRefGoogle ScholarPubMed
Wang, JY, Chambon, C, Lu, CD, Huang, KW, Vivares, CP and Texier, C (2007) A proteomic-based approach for the characterization of some major structural proteins involved in host–parasite relationships from the silkworm parasite Nosema bombycis (microsporidia). Proteomics 7, 1461–1472.CrossRefGoogle ScholarPubMed
Wang, S, Tang, NH, Lara-Gonzalez, P, Zhao, Z, Cheerambathur, DK, Prevo, B, Chisholm, AD, Desai, A and Oegema, K (2017) A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans. Development (Cambridge, England) 144, 2694–2701.Google ScholarPubMed
Weidner, E, Canning, EU, Rutledge, CR and Meek, CL (1999) Mosquito (Diptera: Culicidae) host compatibility and vector competency for the human myositic parasite Trachipleistophora hominis (Phylum Microspora). Journal of Medical Entomology 36, 522–525.CrossRefGoogle ScholarPubMed
Williams, BA, Hirt, RP, Lucocq, JM and Embley, TM (2002) A mitochondrial remnant in the microsporidian Trachipleistophora hominis. Nature 418, 865–869.CrossRefGoogle ScholarPubMed
Zhang, J, Zheng, N and Zhou, P (2003) Exploring the functional complexity of cellular proteins by protein knockout. Proceedings of the National Academy of Sciences of the United States of America 100, 14127–14132.CrossRefGoogle ScholarPubMed
Zheng, S, Huang, Y, Huang, H, Yu, B, Zhou, N, Wei, J, Pan, G, Li, C and Zhou, Z (2021) The role of NbTMP1, a surface protein of sporoplasm, in Nosema bombycis infection. Parasites & Vectors 14, 81.CrossRefGoogle ScholarPubMed