Helical Reconstruction of VP39 Reveals Principles for Baculovirus Nucleocapsid Assembly

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Jan 4

PMID 38177118

Authors

Friederike M C Benning

Simon Jenni

Coby Y Garcia

Tran H Nguyen

Xuewu Zhang

Luke H Chao

Affiliations

Soon will be listed here.

Abstract

Baculoviruses are insect-infecting pathogens with wide applications as biological pesticides, in vitro protein production vehicles and gene therapy tools. Its cylindrical nucleocapsid, which encapsulates and protects the circular double-stranded viral DNA encoding proteins for viral replication and entry, is formed by the highly conserved major capsid protein VP39. The mechanism for VP39 assembly remains unknown. We use electron cryomicroscopy to determine a 3.2 Å helical reconstruction of an infectious nucleocapsid of Autographa californica multiple nucleopolyhedrovirus, revealing how dimers of VP39 assemble into a 14-stranded helical tube. We show that VP39 comprises a distinct protein fold conserved across baculoviruses, which includes a Zinc finger domain and a stabilizing intra-dimer sling. Analysis of sample polymorphism shows that VP39 assembles in several closely-related helical geometries. This VP39 reconstruction reveals general principles for baculoviral nucleocapsid assembly.

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References

Hong M, Li T, Xue W, Zhang S, Cui L, Wang H . Genetic engineering of baculovirus-insect cell system to improve protein production. Front Bioeng Biotechnol. 2022; 10:994743. PMC: 9530357. DOI: 10.3389/fbioe.2022.994743. View

Fang M, Nie Y, Theilmann D . AcMNPV EXON0 (AC141) which is required for the efficient egress of budded virus nucleocapsids interacts with beta-tubulin. Virology. 2009; 385(2):496-504. DOI: 10.1016/j.virol.2008.12.023. View

Grant T, Rohou A, Grigorieff N . TEM, user-friendly software for single-particle image processing. Elife. 2018; 7. PMC: 5854467. DOI: 10.7554/eLife.35383. View

Wang Y, Yu X, Yip C, Strynadka N, Egelman E . Structural polymorphism in bacterial EspA filaments revealed by cryo-EM and an improved approach to helical reconstruction. Structure. 2006; 14(7):1189-96. DOI: 10.1016/j.str.2006.05.018. View

Dohner K, Ramos-Nascimento A, Bialy D, Anderson F, Hickford-Martinez A, Rother F . Importin α1 is required for nuclear import of herpes simplex virus proteins and capsid assembly in fibroblasts and neurons. PLoS Pathog. 2018; 14(1):e1006823. PMC: 5773220. DOI: 10.1371/journal.ppat.1006823. View

Bui K, Yagi T, Yamamoto R, Kamiya R, Ishikawa T . Polarity and asymmetry in the arrangement of dynein and related structures in the Chlamydomonas axoneme. J Cell Biol. 2012; 198(5):913-25. PMC: 3432765. DOI: 10.1083/jcb.201201120. View

Altschul S, Gish W, Miller W, Myers E, Lipman D . Basic local alignment search tool. J Mol Biol. 1990; 215(3):403-10. DOI: 10.1016/S0022-2836(05)80360-2. View

Scheres S . RELION: implementation of a Bayesian approach to cryo-EM structure determination. J Struct Biol. 2012; 180(3):519-30. PMC: 3690530. DOI: 10.1016/j.jsb.2012.09.006. View

Williams C, Headd J, Moriarty N, Prisant M, Videau L, Deis L . MolProbity: More and better reference data for improved all-atom structure validation. Protein Sci. 2017; 27(1):293-315. PMC: 5734394. DOI: 10.1002/pro.3330. View

10.

Scheres S . Amyloid structure determination in RELION-3.1. Acta Crystallogr D Struct Biol. 2020; 76(Pt 2):94-101. PMC: 7008511. DOI: 10.1107/S2059798319016577. View

11.

Danquah J, Botchway S, Jeshtadi A, King L . Direct interaction of baculovirus capsid proteins VP39 and EXON0 with kinesin-1 in insect cells determined by fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy. J Virol. 2011; 86(2):844-53. PMC: 3255796. DOI: 10.1128/JVI.06109-11. View

12.

Egelman E . Reconstruction of helical filaments and tubes. Methods Enzymol. 2010; 482:167-83. PMC: 3245864. DOI: 10.1016/S0076-6879(10)82006-3. View

13.

Winn M, Ballard C, Cowtan K, Dodson E, Emsley P, Evans P . Overview of the CCP4 suite and current developments. Acta Crystallogr D Biol Crystallogr. 2011; 67(Pt 4):235-42. PMC: 3069738. DOI: 10.1107/S0907444910045749. View

14.

Blissard G, Theilmann D . Baculovirus Entry and Egress from Insect Cells. Annu Rev Virol. 2018; 5(1):113-139. DOI: 10.1146/annurev-virology-092917-043356. View

15.

Pearson M, Russell R, Rohrmann G, Beaudreau G . p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location. Virology. 1988; 167(2):407-13. View

16.

Huang W, Tian X, Wu Y, Zhong J, Yu L, Hu S . Suppression of gastric cancer growth by baculovirus vector-mediated transfer of normal epithelial cell specific-1 gene. World J Gastroenterol. 2008; 14(38):5810-5. PMC: 2751889. DOI: 10.3748/wjg.14.5810. View

17.

Bai H, Hu Y, Hu X, Li J, Mu J, Zhou Y . Major capsid protein of Autographa californica multiple nucleopolyhedrovirus contributes to the promoter activity of the very late viral genes. Virus Res. 2019; 273:197758. DOI: 10.1016/j.virusres.2019.197758. View

18.

Wang C, Li F, Yang Y, Guo H, Wu C, Wang S . Recombinant baculovirus containing the diphtheria toxin A gene for malignant glioma therapy. Cancer Res. 2006; 66(11):5798-806. DOI: 10.1158/0008-5472.CAN-05-4514. View

19.

Balani P, Boulaire J, Zhao Y, Zeng J, Lin J, Wang S . High mobility group box2 promoter-controlled suicide gene expression enables targeted glioblastoma treatment. Mol Ther. 2009; 17(6):1003-11. PMC: 2835195. DOI: 10.1038/mt.2009.22. View

20.

Rohou A, Grigorieff N . CTFFIND4: Fast and accurate defocus estimation from electron micrographs. J Struct Biol. 2015; 192(2):216-21. PMC: 6760662. DOI: 10.1016/j.jsb.2015.08.008. View