Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence
Abstract
1. Introduction
2. Methods
2.1. Cells, Viruses, and Insects
2.2. RNA Isolation, Electrophoresis, and Blotting
2.3. Northern Blot Hybridization and Analysis
2.4. Temporal CHIA and V-CATH Protein Analysis
2.5. Chitinase and Cathepsin Activity Assays
3. Results
3.1. Dual Reprogramming of chiA and v-cath Expression
3.2. CHIA and proV-CATH Protein Expression Profiles from Acp6.9-chiA/polh-cath
3.3. Dual Transcriptional Reprogramming Increases CHIA and V-CATH Enzymatic Activity
3.4. Dual chiA/v-cath Reprogramming Does Not Affect Virus Replication
3.5. CHIA and V-CATH Expression in Infected Larvae
3.6. CHIA and V-CATH Enzyme Activity in Infected Larvae
3.7. Dual chiA/v-cath Reprogramming Increases Host Mortality
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Hawtin, R.E.; Zarkowska, T.; Arnold, K.; Thomas, C.J.; Gooday, G.W.; King, L.A.; Kuzio, J.A.; Possee, R.D. Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes. Virology 1997, 238, 243–253. [Google Scholar] [CrossRef]
- O’Reilly, D.R. Auxiliary Genes of Baculoviruses. In The Baculoviruses; Miller, L.K., Ed.; Springer: Boston, MA, USA, 1997. [Google Scholar] [CrossRef]
- Corrado, G.; Arciello, S.; Fanti, P.; Fiandra, L.; Garonna, A.; Digilio, M.C.; Lorito, M.; Giordana, B.; Pennacchio, F.; Rao, R. The Chitinase A from the baculovirus AcMNPV enhances resistance to both fungi and herbivorous pests in tobacco. Transgenic Res. 2008, 17, 557–571. [Google Scholar] [CrossRef] [PubMed]
- Lima, A.A.; Aragao, C.W.; de Castro, M.E.; Oliveira, J.V.; Sosa Gomez, D.R.; Ribeiro, B.M. A recombinant Anticarsia gemmatalis MNPV harboring chiA and v-cath genes from Choristoneura fumiferana defective NPV induce host liquefaction and increased insecticidal activity. PLoS ONE 2013, 8, e74592. [Google Scholar] [CrossRef] [PubMed]
- Wang, Y.; Choi, J.Y.; Roh, J.Y.; Tao, X.Y.; Liu, Q.; Lee, J.H.; Kim, J.S.; Kim, W.J.; Je, Y.H. Insecticidal activity of the chitinase from the Spodoptera litura nucleopolyhedrovirus. Entomol. Res. 2013, 43, 63–69. [Google Scholar] [CrossRef]
- Saville, G.P.; Patmanidi, A.L.; Possee, R.D.; King, L.A. Deletion of the Autographa californica nucleopolyhedrovirus chitinase KDEL motif and in vitro and in vivo analysis of the modified virus. J. Gen. Virol. 2004, 85, 821–831. [Google Scholar] [CrossRef] [PubMed]
- Saville, G.P.; Thomas, C.J.; Possee, R.D.; King, L.A. Partial redistribution of the Autographa californica nucleopolyhedrovirus chitinase in virus-infected cells accompanies mutation of the carboxy-terminal KDEL ER-retention motif. J. Gen. Virol. 2002, 83, 685–694. [Google Scholar] [CrossRef]
- Gopalakrishnan, B.; Muthukrishnan, S.; Kramer, K.J. Baculovirus-Mediated Expression of a Manduca-Sexta Chitinase Gene-Properties of the Recombinant Protein. Insect Biochem. Molec. 1995, 25, 255–265. [Google Scholar] [CrossRef]
- Li, H.R.; Tang, H.L.; Sivakumar, S.; Philip, J.; Harrison, R.L.; Gatehouse, J.A.; Bonning, B.C. Insecticidal activity of a basement membrane-degrading protease against Heliothis virescens (Fabricius) and Acyrthosiphon pisum (Harris). J. Insect Physiol. 2008, 54, 777–789. [Google Scholar] [CrossRef]
- Slack, J.M.; Ribeiro, B.M.; de Souza, M.L. The gp64 locus of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus contains a 3′ repair exonuclease homologue and lacks v-cath and ChiA genes. J. Gen. Virol. 2004, 85, 211–219. [Google Scholar] [CrossRef]
- Hodgson, J.J.; Arif, B.M.; Krell, P.J. Interaction of Autographa californica multiple nucleopolyhedrovirus cathepsin protease progenitor (proV-CATH) with insect baculovirus chitinase as a mechanism for proV-CATH cellular retention. J. Virol. 2011, 85, 3918–3929. [Google Scholar] [CrossRef]
- Hom, L.G.; Ohkawa, T.; Trudeau, D.; Volkman, L.E. Autographa californica M nucleopolyhedrovirus ProV-CATH is activated during infected cell death. Virology 2002, 296, 212–218. [Google Scholar] [CrossRef] [PubMed][Green Version]
- Hodgson, J.J.; Krell, P.J.; Passarelli, A.L. Mature viral cathepsin is required for release of viral occlusion bodies from Autographa californica multiple nucleopolyhedrovirus-infected cells. Virology 2021, 556, 23–32. [Google Scholar] [CrossRef]
- Daimon, T.; Katsuma, S.; Shimada, T. Mutational analysis of active site residues of chitinase from Bombyx mori nucleopolyhedrovirus. Virus Res. 2007, 124, 168–175. [Google Scholar] [CrossRef]
- Hom, L.G.; Volkman, L.E. Autographa californica M nucleopolyhedrovirus chiA is required for processing of V-CATH. Virology 2000, 277, 178–183. [Google Scholar] [CrossRef] [PubMed][Green Version]
- Katsuma, S.; Nakanishi, T.; Daimon, T.; Shimada, T. N-linked glycans located in the pro-region of Bombyx mori nucleopolyhedrovirus V-CATH are essential for the proper folding of V-CATH and V-CHIA. J. Gen. Virol. 2009, 90, 170–176. [Google Scholar] [CrossRef]
- Hodgson, J.J.; Arif, B.M.; Krell, P.J. Role of interactions between Autographa californica multiple nucleopolyhedrovirus procathepsin and chitinase chitin-binding or active-site domains in viral cathepsin processing. J. Virol. 2013, 87, 3471–3483. [Google Scholar] [CrossRef] [PubMed][Green Version]
- Ishimwe, E.; Hodgson, J.J.; Clem, R.J.; Passarelli, A.L. Reaching the melting point: Degradative enzymes and protease inhibitors involved in baculovirus infection and dissemination. Virology 2015, 479, 637–649. [Google Scholar] [CrossRef] [PubMed]
- Hodgson, J.J.; Arif, B.M.; Krell, P.J. Reprogramming the chiA expression profile of Autographa californica multiple nucleopolyhedrovirus. J. Gen. Virol. 2007, 88, 2479–2487. [Google Scholar] [CrossRef] [PubMed]
- Hodgson, J.J.; Arif, B.M.; Krell, P.J. Autographa californica multiple nucleopolyhedrovirus and Choristoneura fumiferana multiple nucleopolyhedrovirus v-cath genes are expressed as pre-proenzymes. J. Gen. Virol. 2009, 90, 995–1000. [Google Scholar] [CrossRef] [PubMed]
- O’Reilly, D.R. Use of baculovirus expression vectors. Methods Mol. Biol. 1997, 62, 235–246. [Google Scholar] [CrossRef]
- Hill, J.E.; Kuzio, J.; Faulkner, P. Identification and characterization of the v-cath gene of the baculovirus, CfMNPV. Gene Struct. Expr. 1995, 1264, 275–278. [Google Scholar] [CrossRef] [PubMed]
- Daimon, T.; Katsuma, S.; Kang, W.K.; Shimada, T. Functional characterization of chitinase from Cydia pomonella granulovirus. Arch. Virol. 2007, 152, 1655–1664. [Google Scholar] [CrossRef]
- Katsuma, S.; Nakanishi, T.; Shimada, T. Bombyx mori nucleopolyhedrovirus FP25K is essential for maintaining a steady-state level of v-cath expression throughout the infection. Virus Res. 2009, 140, 155–160. [Google Scholar] [CrossRef] [PubMed]
- Slack, J.M.; Kuzio, J.; Faulkner, P. Characterization of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J. Gen. Virol. 1995, 76 Pt 5, 1091–1098. [Google Scholar] [CrossRef]
- Hom, L.G.; Volkman, L.E. Preventing proteolytic artifacts in the baculovirus expression system. Biotechniques 1998, 25, 18–20. [Google Scholar] [CrossRef] [PubMed]
- Ishimwe, E.; Hodgson, J.J.; Passarelli, A.L. Expression of the Cydia pomonella granulovirus matrix metalloprotease enhances Autographa californica multiple nucleopolyhedrovirus virulence and can partially substitute for viral cathepsin. Virology 2015, 481, 166–178. [Google Scholar] [CrossRef] [PubMed]
- Friesen, P.D.; Miller, L.K. Temporal regulation of baculovirus RNA: Overlapping early and late transcripts. J. Virol. 1985, 54, 392–400. [Google Scholar] [CrossRef]
- Ooi, B.G.; Rankin, C.; Miller, L.K. Downstream sequences augment transcription from the essential initiation site of a baculovirus polyhedrin gene. J. Mol. Biol. 1989, 210, 721–736. [Google Scholar] [CrossRef]
- Possee, R.D.; Howard, S.C. Analysis of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Res. 1987, 15, 10233–10248. [Google Scholar] [CrossRef]
- Hawtin, R.E.; Arnold, K.; Ayres, M.D.; Zanotto, P.M.; Howard, S.C.; Gooday, G.W.; Chappell, L.H.; Kitts, P.A.; King, L.A.; Possee, R.D. Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 1995, 212, 673–685. [Google Scholar] [CrossRef]
- Ohkawa, T.; Majima, K.; Maeda, S. A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. J. Virol. 1994, 68, 6619–6625. [Google Scholar] [CrossRef] [PubMed]
- Moscardi, F.; de Souza, M.L.; de Castro, M.E.B.; Moscardi, M.L.; Szewczyk, B. Baculovirus Pesticides: Present State and Future Perspectives. In Microbes and Microbial Technology: Agricultural and Environmental Applications; Springer: New York, NY, USA, 2011; pp. 415–445. [Google Scholar] [CrossRef]
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Hodgson, J.J.; Passarelli, A.L.; Krell, P.J. Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence. Viruses 2023, 15, 503. https://doi.org/10.3390/v15020503
Hodgson JJ, Passarelli AL, Krell PJ. Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence. Viruses. 2023; 15(2):503. https://doi.org/10.3390/v15020503
Chicago/Turabian StyleHodgson, Jeffrey J., A. Lorena Passarelli, and Peter J. Krell. 2023. "Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence" Viruses 15, no. 2: 503. https://doi.org/10.3390/v15020503
APA StyleHodgson, J. J., Passarelli, A. L., & Krell, P. J. (2023). Transcriptional Reprogramming of Autographa Californica Multiple Nucleopolyhedrovirus Chitinase and Cathepsin Genes Enhances Virulence. Viruses, 15(2), 503. https://doi.org/10.3390/v15020503