Dendrolimus punctatus causes great damage to pine forests worldwide. Dendrolimus punctatus cypovirus 1 (DpCPV-1) is an important pathogen of D. punctatus. However, the mechanism of DpCPV-1 cell entry has not been elucidated. In this study, we revealed that both GTase and MTase domains of VP3 (B-spike) and VP4 (A-spike) of DpCPV-1 interacted with the midgut proteins of Bombyx mori. Binding and competition assays revealed that GTase, MTase and VP4 played roles as viral attachment proteins. Far-Western blotting and LC-MS/MS analyses identified that heat shock protein 70 (BmHSP70), glutamate dehydrogenase (BmGDH), and angiotensin-converting enzyme (BmACE) in the midgut proteins as ligand candidates of the viral attachment proteins, and this was further verified by co-immunoprecipitation and fluorescence co-localization assays. Viral binding to the host midgut in vitro was inhibited by pre-treating B. mori midgut proteins with anti-BmHSP70, anti-BmGDH, anti-BmACE antibodies singly and in combination. Incubating DpCPV-1 virions with prokaryotically expressed BmHSP70, BmGDH, and BmACE also decreased viral attachment to the host midgut. In vivo bioassays revealed that viral infection in Helicoverpa armigera was partially neutralized by BmHSP70, BmGDH, and BmACE. Taking together, we concluded that HSP70, GDH, and ACE mediate DpCPV attachment and entry via binding to the viral attachment proteins, VP3 and VP4. The findings provide foundation for further understanding the entry mechanisms of cypoviruses.
YuX, GeP, JiangJ, AtanasovI, ZhouZH. Atomic model of CPV reveals the mechanism used by this single-shelled virus to economically carry out functions conserved in multishelled reoviruses. Structure2011; 19:652–661 [View Article] [PubMed]
ChengC, ShaoY, SuL, ZhouY, SunX. Interactions among Dendrolimus punctatus cypovirus proteins and identification of the genomic segment encoding its A-spike. J Gen Virol2014; 95:1532–1538 [View Article] [PubMed]
XuG, WilsonW, MechamJ, MurphyK, ZhouEM et al. VP7: an attachment protein of bluetongue virus for cellular receptors in Culicoides variipennis. J Gen Virol1997; 78 (Pt 7):1617–1623 [View Article] [PubMed]
LudertJE, FengN, YuJH, BroomeRL, HoshinoY et al. Genetic mapping indicates that VP4 is the rotavirus cell attachment protein in vitro and in vivo. J Virol1996; 70:487–493 [View Article] [PubMed]
GuglielmiKM, JohnsonEM, StehleT, DermodyTS. Attachment and cell entry of mammalian orthoreovirus. In RoyP. eds Reoviruses: Entry, Assembly and Morphogenesis (Current Topics in Microbiology and Immunology)2006 pp 1–38
RothAN, AravamudhanP, Fernández de CastroI, TenorioR, RiscoC et al. Ins and outs of reovirus: vesicular trafficking in viral entry and egress. Trends Microbiol2021; 29:363–375 [View Article] [PubMed]
KirchnerE, GuglielmiKM, StraussHM, DermodyTS, StehleT et al. Structure of reovirus σ1 in complex with its receptor junctional adhesion molecule-A. PLoS Pathog2008; 4:e1000235 [View Article]
Torres-FloresJM, Silva-AyalaD, EspinozaMA, LópezS, AriasCF. The tight junction protein JAM-A functions as coreceptor for rotavirus entry into MA104 cells. Virology2015; 475:172–178 [View Article] [PubMed]
ZhangY, CaoG, ZhuL, ChenF, ZarMS et al. Integrin beta and receptor for activated protein kinase C are involved in the cell entry of Bombyx mori cypovirus. Appl Microbiol Biotechnol2017; 101:3703–3716 [View Article] [PubMed]
Hai-menK, Fu-yanW, Zhong-dongY, Hai-yanC, Xiao-manLI et al. Protection of mice with DNA vaccines encoding influenza virus matrix protein. Life Science Research2009; 13:42–45
ZhouY, QinT, XiaoY, QinF, LeiC et al. Genomic and biological characterization of a new cypovirus isolated from Dendrolimus punctatus. PLoS ONE2014; 9:e113201 [View Article]
KalbfleischJ, PrenticeR. Competing risks and multistate models. In The Statistical Analysis of Failure Time Data John Wiley & Sons, Inc; 2011 pp 247–277
HongJ, DuanJ, ZhaoS, PengH. The sensitivity of Bombyx mori larvae to Dendrolimus punctatus cytoplasmic polyhedrosis virus. Acta Entomologica Sinica2003; 46:409–416
LiW, MooreMJ, VasilievaN, SuiJ, WongSK et al. Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature2003; 426:450–454 [View Article] [PubMed]
Reyes-Del ValleJ, Chávez-SalinasS, MedinaF, Del AngelRM. Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells. J Virol2005; 79:4557–4567 [View Article] [PubMed]
DasS, LaxminarayanaSV, ChandraN, RaviV, DesaiA. Heat shock protein 70 on Neuro2a cells is a putative receptor for Japanese encephalitis virus. Virology2009; 385:47–57 [View Article] [PubMed]
WangXJ. Heat-shock protein 70 is associated with the entry of Marek’s disease virus into fibroblast. Acta Virol2011; 55:189–194 [View Article] [PubMed]
SurteesR, DowallSD, ShawA, ArmstrongS, HewsonR et al. Heat shock protein 70 family members interact with Crimean-Congo hemorrhagic fever virus and hazara virus nucleocapsid proteins and perform a functional role in the nairovirus replication cycle. J Virol2016; 90:9305–9316 [View Article] [PubMed]
ZhaoD, HanK, LiuQ, HuangX, YangJ et al. Identification of Tembusu virus receptor protein in the duck embryo fibroblasts. Journal of Southern Argiculture2019; 50:173–178
WangY, ZhangH, MaD, DengX, WuD et al. Hsp70 is a potential therapeutic target for echovirus 9 infection. Front Mol Biosci2020; 7:146 [View Article] [PubMed]
PlaitakisA, ZaganasI. Regulation of human glutamate dehydrogenases: implications for glutamate, ammonia and energy metabolism in brain. J Neurosci Res2001; 66:899–908 [View Article] [PubMed]
JoanisseDR, StoreyKB. Mitochondrial enzymes during overwintering in two species of cold-hardy gall insects. Insect Biochem Mol Biol1994; 24:145–150 [View Article]
LiuZ-F, LiangY-Y, SunX-T, YangJ, ZhangP-J et al. Analysis of differentially expressed genes of Chrysoperla sinica related to flight capacity by transcriptome. J Insect Sci2021; 21:18. [View Article] [PubMed]
LamangoNS, NachmanRJ, HayesTK, StreyA, IsaacRE. Hydrolysis of insect neuropeptides by an angiotensin-converting enzyme from the housefly, Musca domestica. Peptides1997; 18:47–52 [View Article] [PubMed]
IsaacRE, SchoofsL, WilliamsTA, CorvolP, VeelaertD et al. Toward a role for angiotensin-converting enzyme in insects. Annals NY Acad Sci1998; 839:288–292 [View Article]