Mammarenavirus Z Protein Myristoylation and Oligomerization Are Not Required for Its Dose-Dependent Inhibitory Effect on vRNP Activity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Line and Compounds
2.2. Plasmids
2.3. Minigenome Assay
2.4. VLP Assay
2.5. Immunoblotting
2.6. Immunofluorescence Imaging
3. Results
3.1. Effect of NMTi on Z-Mediated Inhibition of vRNP Activity
3.2. Role of N-Myristoylation on Z-Mediated Inhibition of vRNP Activity
3.3. Role of Z Oligomerization on Z-Mediated Inhibition of vRNP Activity and Z Budding Activity
4. Discussion
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
- Yadav, K.; Mathur, G.; Ford, B.; Miller, R.; Group, C.W. A Case Cluster of Lymphocytic Choriomeningitis Virus Transmitted Via Organ Transplantation.: Abstract# D2381. Transplantation 2014, 98, 768. [Google Scholar]
- Sayyad, L.E.; Smith, K.L.; Sadigh, K.S.; Cossaboom, C.M.; Choi, M.J.; Whitmer, S.; Cannon, D.; Krapiunaya, I.; Morales-Betoulle, M.; Annambhotla, P.; et al. Severe Non–Donor-Derived Lymphocytic Choriomeningitis Virus Infection in 2 Solid Organ Transplant Recipients. Open Forum Infect. Dis. 2025, 12, ofaf002. [Google Scholar] [CrossRef]
- Schafer, I.J.; Miller, R.; Ströher, U.; Knust, B.; Nichol, S.T.; Rollin, P.E.; Centers for Disease Control and Prevention (CDC). Notes from the Field: A Cluster of Lymphocytic Choriomeningitis Virus Infections Transmitted Through Organ Transplantation—Iowa, 2013. MMWR Morb. Mortal. Wkly. Rep. 2014, 63, 249. [Google Scholar] [CrossRef]
- Carrillo-Bustamante, P.; Nguyen, T.H.T.; Oestereich, L.; Günther, S.; Guedj, J.; Graw, F. Determining Ribavirin’s Mechanism of Action against Lassa Virus Infection. Sci. Rep. 2017, 7, 11693. [Google Scholar] [CrossRef]
- Radoshitzky, S.R.; Buchmeier, M.; de la Torre, J.C. Emerging Viruses: Arenaviridae. In Fields Virology, 7th ed.; Knipe, D., Howley, P., Whelan, S., Eds.; Wolters Kluwer: Philadelphia, PA, USA, 2021; Volume I, ISBN 978-1-975112-54-7. [Google Scholar]
- Kunz, S.; Edelmann, K.H.; de la Torre, J.-C.; Gorney, R.; Oldstone, M.B.A. Mechanisms for Lymphocytic Choriomeningitis Virus Glycoprotein Cleavage, Transport, and Incorporation into Virions. Virology 2003, 314, 168–178. [Google Scholar] [CrossRef]
- Rojek, J.M.; Lee, A.M.; Nguyen, N.; Spiropoulou, C.F.; Kunz, S. Site 1 Protease Is Required for Proteolytic Processing of the Glycoproteins of the South American Hemorrhagic Fever Viruses Junin, Machupo, and Guanarito. J. Virol. 2008, 82, 6045–6051. [Google Scholar] [CrossRef]
- Fedeli, C.; Moreno, H.; Kunz, S. Novel Insights into Cell Entry of Emerging Human Pathogenic Arenaviruses. J. Mol. Biol. 2018, 430, 1839–1852. [Google Scholar] [CrossRef]
- York, J.; Nunberg, J.H. Role of the Stable Signal Peptide of Junín Arenavirus Envelope Glycoprotein in pH-Dependent Membrane Fusion. J. Virol. 2006, 80, 7775–7780. [Google Scholar] [CrossRef]
- York, J.; Nunberg, J.H. Myristoylation of the Arenavirus Envelope Glycoprotein Stable Signal Peptide Is Critical for Membrane Fusion but Dispensable for Virion Morphogenesis. J. Virol. 2016, 90, 8341–8350. [Google Scholar] [CrossRef]
- Perez, M.; Greenwald, D.L.; de La Torre, J.C. Myristoylation of the RING Finger Z Protein Is Essential for Arenavirus Budding. J. Virol. 2004, 78, 11443–11448. [Google Scholar] [CrossRef]
- Capul, A.A.; Perez, M.; Burke, E.; Kunz, S.; Buchmeier, M.J.; de la Torre, J.C. Arenavirus Z-Glycoprotein Association Requires Z Myristoylation but Not Functional RING or Late Domains. J. Virol. 2007, 81, 9451–9460. [Google Scholar] [CrossRef]
- Cordo, S.M.; Candurra, N.A.; Damonte, E.B. Myristic Acid Analogs Are Inhibitors of Junin Virus Replication. Microbes Infect. 1999, 1, 609–614. [Google Scholar] [CrossRef]
- Kallemeijn, W.W.; Lueg, G.A.; Faronato, M.; Hadavizadeh, K.; Grocin, A.G.; Song, O.-R.; Howell, M.; Calado, D.P.; Tate, E.W. Validation and Invalidation of Chemical Probes for the Human N-Myristoyltransferases. Cell Chem. Biol. 2019, 26, 892–900.e4. [Google Scholar] [CrossRef]
- Ramljak, I.C.; Stanger, J.; Real-Hohn, A.; Dreier, D.; Wimmer, L.; Redlberger-Fritz, M.; Fischl, W.; Klingel, K.; Mihovilovic, M.D.; Blaas, D.; et al. Cellular N-Myristoyltransferases Play a Crucial Picornavirus Genus-Specific Role in Viral Assembly, Virion Maturation, and Infectivity. PLoS Pathog. 2018, 14, e1007203. [Google Scholar] [CrossRef]
- Witwit, H.; Betancourt, C.A.; Cubitt, B.; Khafaji, R.; Kowalski, H.; Jackson, N.; Ye, C.; Martinez-Sobrido, L.; de la Torre, J.C. Cellular N-Myristoyl Transferases Are Required for Mammarenavirus Multiplication. Viruses 2024, 16, 1362. [Google Scholar] [CrossRef]
- Cornu, T.I.; de la Torre, J.C. Characterization of the Arenavirus RING Finger Z Protein Regions Required for Z-Mediated Inhibition of Viral RNA Synthesis. J. Virol. 2002, 76, 6678–6688. [Google Scholar] [CrossRef]
- Jácamo, R.; López, N.; Wilda, M.; Franze-Fernández, M.T. Tacaribe Virus Z Protein Interacts with the L Polymerase Protein To Inhibit Viral RNA Synthesis. J. Virol. 2003, 77, 10383–10393. [Google Scholar] [CrossRef]
- López, N.; Jácamo, R.; Franze-Fernández, M.T. Transcription and RNA Replication of Tacaribe Virus Genome and Antigenome Analogs Require N and L Proteins: Z Protein Is an Inhibitor of These Processes. J. Virol. 2001, 75, 12241–12251. [Google Scholar] [CrossRef]
- Liu, L.; Wang, P.; Liu, A.; Zhang, L.; Yan, L.; Guo, Y.; Xiao, G.; Rao, Z.; Lou, Z. Structure Basis for Allosteric Regulation of Lymphocytic Choriomeningitis Virus Polymerase Function by Z Matrix Protein. Protein Cell 2023, 14, 703–707. [Google Scholar] [CrossRef]
- Cornu, T.I.; de la Torre, J.C. RING Finger Z Protein of Lymphocytic Choriomeningitis Virus (LCMV) Inhibits Transcription and RNA Replication of an LCMV S-Segment Minigenome. J. Virol. 2001, 75, 9415–9426. [Google Scholar] [CrossRef]
- Iwasaki, M.; de la Torre, J.C. A Highly Conserved Leucine in Mammarenavirus Matrix Z Protein Is Required for Z Interaction with the Virus L Polymerase and Z Stability in Cells Harboring an Active Viral Ribonucleoprotein. J. Virol. 2018, 92, e02256-17. [Google Scholar] [CrossRef] [PubMed]
- Hastie, K.M.; Zandonatti, M.; Liu, T.; Li, S.; Woods, V.L.; Saphire, E.O. Crystal Structure of the Oligomeric Form of Lassa Virus Matrix Protein Z. J. Virol. 2016, 90, 4556–4562. [Google Scholar] [CrossRef] [PubMed]
- Loureiro, M.E.; Wilda, M.; Levingston Macleod, J.M.; D’Antuono, A.; Foscaldi, S.; Buslje, C.M.; Lopez, N. Molecular Determinants of Arenavirus Z Protein Homo-Oligomerization and L Polymerase Binding. J. Virol. 2011, 85, 12304–12314. [Google Scholar] [CrossRef] [PubMed]
- Qiu, W.; Hutchinson, A.; Wernimont, A.; Lin, Y.-H.; Kania, A.; Ravichandran, M.; Kozieradzki, I.; Cossar, D.; Schapira, M.; Arrowsmith, C.H.; et al. Crystal Structure of Human Type-I N-Myristoyltransferase with Bound Myristoyl-CoA and Inhibitor DDD85646. 2010; To be published. [Google Scholar] [CrossRef]
- Volpon, L.; Osborne, M.J.; Capul, A.A.; de la Torre, J.C.; Borden, K.L.B. Structural Characterization of the Z RING-eIF4E Complex Reveals a Distinct Mode of Control for eIF4E. Proc. Natl. Acad. Sci. USA 2010, 107, 5441–5446. [Google Scholar] [CrossRef]
- Meng, E.C.; Goddard, T.D.; Pettersen, E.F.; Couch, G.S.; Pearson, Z.J.; Morris, J.H.; Ferrin, T.E. UCSF ChimeraX: Tools for Structure Building and Analysis. Protein Sci. 2023, 32, e4792. [Google Scholar] [CrossRef]
- Perez, M.; Craven, R.C.; De La Torre, J.C. The Small RING Finger Protein Z Drives Arenavirus Budding: Implications for Antiviral Strategies. Proc. Natl. Acad. Sci. USA 2003, 100, 12978–12983. [Google Scholar] [CrossRef]
- Baudin, F.; Petit, I.; Weissenhorn, W.; Ruigrok, R.W. In Vitro Dissection of the Membrane and RNP Binding Activities of Influenza Virus M1 Protein. Virology 2001, 281, 102–108. [Google Scholar] [CrossRef]
- Cornu, T.I.; Feldmann, H.; de la Torre, J.C. Cells Expressing the RING Finger Z Protein Are Resistant to Arenavirus Infection. J. Virol. 2004, 78, 2979–2983. [Google Scholar] [CrossRef]
- Kranzusch, P.J.; Whelan, S.P.J. Arenavirus Z Protein Controls Viral RNA Synthesis by Locking a Polymerase–Promoter Complex. Proc. Natl. Acad. Sci. USA 2011, 108, 19743–19748. [Google Scholar] [CrossRef]
- Witwit, H.; Cubitt, B.; Khafaji, R.; Castro, E.M.; Goicoechea, M.; Lorenzo, M.M.; Blasco, R.; Martinez-Sobrido, L.; de la Torre, J.C. Repurposing Drugs for Synergistic Combination Therapies to Counteract Monkeypox Virus Tecovirimat Resistance. Viruses 2025, 17, 92. [Google Scholar] [CrossRef]
- Sangha, R.; Davies, N.M.; Namdar, A.; Chu, M.; Spratlin, J.; Beauchamp, E.; Berthiaume, L.G.; Mackey, J.R. Novel, First-in-Human, Oral PCLX-001 Treatment in a Patient with Relapsed Diffuse Large B-Cell Lymphoma. Curr. Oncol. 2022, 29, 1939–1946. [Google Scholar] [CrossRef] [PubMed]
- Sangha, R.S.; Jamal, R.; Spratlin, J.L.; Kuruvilla, J.; Sehn, L.H.; Weickert, M.; Berthiaume, L.G.; Mackey, J.R. A First-in-Human, Open-Label, Phase I Trial of Daily Oral PCLX-001, an NMT Inhibitor, in Patients with Relapsed/Refractory B-Cell Lymphomas and Advanced Solid Tumors. JCO 2023, 41, e15094. [Google Scholar] [CrossRef]
- Priyamvada, L.; Kallemeijn, W.W.; Faronato, M.; Wilkins, K.; Goldsmith, C.S.; Cotter, C.A.; Ojeda, S.; Solari, R.; Moss, B.; Tate, E.W.; et al. Inhibition of Vaccinia Virus L1 N-Myristoylation by the Host N-Myristoyltransferase Inhibitor IMP-1088 Generates Non-Infectious Virions Defective in Cell Entry. PLoS Pathog. 2022, 18, e1010662. [Google Scholar] [CrossRef] [PubMed]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Witwit, H.; de la Torre, J.C. Mammarenavirus Z Protein Myristoylation and Oligomerization Are Not Required for Its Dose-Dependent Inhibitory Effect on vRNP Activity. BioChem 2025, 5, 10. https://doi.org/10.3390/biochem5020010
Witwit H, de la Torre JC. Mammarenavirus Z Protein Myristoylation and Oligomerization Are Not Required for Its Dose-Dependent Inhibitory Effect on vRNP Activity. BioChem. 2025; 5(2):10. https://doi.org/10.3390/biochem5020010
Chicago/Turabian StyleWitwit, Haydar, and Juan C. de la Torre. 2025. "Mammarenavirus Z Protein Myristoylation and Oligomerization Are Not Required for Its Dose-Dependent Inhibitory Effect on vRNP Activity" BioChem 5, no. 2: 10. https://doi.org/10.3390/biochem5020010
APA StyleWitwit, H., & de la Torre, J. C. (2025). Mammarenavirus Z Protein Myristoylation and Oligomerization Are Not Required for Its Dose-Dependent Inhibitory Effect on vRNP Activity. BioChem, 5(2), 10. https://doi.org/10.3390/biochem5020010