Aufbau von Replikonsystemen für Altwelt-Arenaviren und Herstellung rekombinanter Mopeia-Viren

Abstract

Lassa virus (LASV) causing hemorrhagic fever, Mopeia virus (MOPV) and lymphocytic choriomeningitis virus (LCMV) are the main representatives of the Old World arenaviruses. Minireplicon systems for MOPV and LCMV were established in analogy to the existing LASV system. The functional and physical integrity of replication complexes was tested by luciferase assay, Northern blotting, and coimmunoprecipitation studies. The minigenomes, NPs, and L proteins of LASV and MOPV could be exchanged without loss of function LASV and MOPV L proteins are functional also with LCMV NP, while the LCMV L protein requires homologous NP for activity. Analysis of LASV/LCMV NP chimeras identified a single LCMV-specific NP residue (Ile-53) and the C terminus of NP as being essential for LCMV L protein function. The defect of LASV and MOPV NP in supporting transcriptional activity of LCMV L protein was not caused by a defect in physical NP-L protein interaction. Furthermore the first reverse genetic system for production of infectious MOPV was established. Overall, new reverse genetic systems were established to study the replication cycle of Old World arenaviruses.