title: A Single Amino Acid Change in the Marburg Virus Matrix Protein VP40 Provides a Replicative Advantage in a Species-Specific Manner.
abstract: Marburg virus (MARV) induces severe hemorrhagic fever in humans and nonhuman primates but only transient nonlethal disease in rodents. However, sequential passages of MARV in rodents boosts infection leading to lethal disease. Guinea pig-adapted MARV contains one mutation in the viral matrix protein VP40 at position 184 (VP40D184N). The contribution of the D184N mutation to the efficacy of replication in a new host is unknown. In the present study, we demonstrated that recombinant MARV containing the D184N mutation in VP40 [rMARVVP40(D184N)] grew to higher titers than wild-type recombinant MARV (rMARVWT) in guinea pig cells. Moreover, rMARVVP40(D184N) displayed higher infectivity in guinea pig cells. Comparative analysis of VP40 functions indicated that neither the interferon (IFN)-antagonistic function nor the membrane binding capabilities of VP40 were affected by the D184N mutation. However, the production of VP40-induced virus-like particles (VLPs) and the recruitment of other viral proteins to the budding site was improved by the D184N mutation in guinea pig cells, which resulted in the higher infectivity of VP40D184N-induced infectious VLPs (iVLPs) compared to that of VP40-induced iVLPs. In addition, the function of VP40 in suppressing viral RNA synthesis was influenced by the D184N mutation specifically in guinea pig cells, thus allowing greater rates of transcription and replication. Our results showed that the improved viral fitness of rMARVVP40(D184N) in guinea pig cells was due to the better viral assembly function of VP40D184N and its lower inhibitory effect on viral transcription and replication rather than modulation of the VP40-mediated suppression of IFN signaling.
  
  
  OBJECTIVE
  The increased virulence achieved by virus passaging in a new host was accompanied by mutations in the viral genome. Analyzing how these mutations affect the functions of viral proteins and the ability of the virus to grow within new host cells helps in the understanding of the molecular mechanisms increasing virulence. Using a reverse genetics approach, we demonstrated that a single mutation in MARV VP40 detected in a guinea pig-adapted MARV provided a replicative advantage of rMARVVP40(D184N) in guinea pig cells. Our studies show that this replicative advantage of rMARV VP40D184N was based on the improved functions of VP40 in iVLP assembly and in the regulation of transcription and replication rather than on the ability of VP40 to combat the host innate immunity.
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Koehler
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Larissa
authors.initials: L
authors.lastname: Kolesnikova
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Ulla
authors.initials: U
authors.lastname: Welzel
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Gordian
authors.initials: G
authors.lastname: Schudt
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Herwig
authors.affiliation: Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany German Center for Infection Research (DZIF), Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany becker@staff.uni-marburg.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
mesh_terms: D000220:Adaptation, Biological; D000818:Animals; D002460:Cell Line; D006168:Guinea Pigs; D006801:Humans; D029024:Marburgvirus / Q000235:genetics / Q000502:physiology*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000378:metabolism*; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D050505:Mutant Proteins; C080981:VP40 protein, virus; D014763:Viral Matrix Proteins