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D471G mutation in LCMV-NP affects its ability to self-associate and results in a dominant negative effect in viral
Emilio Ortiz-Riaño1, Benson Y H Cheng, Juan C de la Torre
1Department of Microbiology and Immunology, University of Rochester, Rochester, New York 14642, USA. eortizriano@urmc.rochester.edu
Abstract:
Arenaviruses merit significant interest because several family members are etiological agents of severe hemorrhagic fevers, representing a major burden to public health. Currently, there are no FDA-licensed vaccines against arenaviruses and the only available antiviral therapy is limited to the use of ribavirin that is partially effective. Arenavirus nucleoprotein (NP) is found associated with the genomic RNA forming the viral ribonucleoproteins (vRNPs) that together with the polymerase (L) direct viral replication and transcription. Virion formation requires the recruitment of vRNPs into budding sites, a process in which the arenavirus matrix-like protein (Z) plays a major role. Therefore, proper NP-NP and NP-Z interactions are required for the generation of infectious progeny. In this work we demonstrate the role of the amino acid residue D471 in the self-association of lymphocytic choriomeningitis virus nucleoprotein (LCMV-NP). Amino acid substitutions at this position abrogate NP oligomerization, affecting its ability to mediate replication and transcription of a minigenome reporter plasmid. However, its ability to interact with the Z protein, counteract the cellular interferon response and bind to dsRNA analogs was retained. Additionally, we also document the dominant negative effect of D471G mutation on viral infection, suggesting that NP self-association is an excellent target for the development of new antivirals against arenaviruses.
Insights
Targeting lymphocytic choriomeningitis virus nucleoprotein (LCMV-NP) self-association is a promising antiviral strategy. Mutations disrupting LCMV-NP oligomerization inhibit viral replication while preserving essential functions, offering a potential therapeutic avenue.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Arenaviruses cause severe hemorrhagic fevers, posing a significant public health threat.
- Current treatments for arenavirus infections are limited, with no FDA-approved vaccines and partially effective antiviral drugs like ribavirin.
Purpose of the Study:
- To investigate the role of specific amino acid residues in the self-association of lymphocytic choriomeningitis virus nucleoprotein (LCMV-NP).
- To evaluate the impact of disrupting LCMV-NP self-association on viral replication and potential as an antiviral target.
Main Methods:
- Site-directed mutagenesis was used to alter the D471 residue in LCMV-NP.
- Assays were performed to assess NP oligomerization, minigenome replication and transcription, NP-Z protein interaction, and dsRNA binding.
- The effect of the D471G mutation on viral infection was evaluated using a dominant-negative approach.
Main Results:
- The D471 residue is crucial for LCMV-NP self-association; substitutions at this position abrogate NP oligomerization.
- Disruption of NP oligomerization impaired its ability to mediate viral replication and transcription.
- Importantly, the D471G mutation retained NP's ability to interact with the Z protein, counteract interferon responses, and bind dsRNA.
Conclusions:
- LCMV-NP self-association is essential for viral replication and represents a viable target for novel antiviral development.
- Targeting NP oligomerization offers a potential strategy for developing new therapies against arenavirus infections.
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