title: The marburg virus 3' noncoding region structurally and functionally differs from that of ebola virus.
abstract: We have previously shown that the first transcription start signal (TSS) of Zaire Ebola virus (ZEBOV) is involved in formation of an RNA secondary structure regulating VP30-dependent transcription activation. Interestingly, transcription of Marburg virus (MARV) minigenomes occurs independently of VP30. In this study, we analyzed the structure of the MARV 3' noncoding region and its influence on VP30 necessity. Secondary structure formation of the TSS of the first gene was experimentally determined and showed substantial differences from the structure formed by the ZEBOV TSS. Chimeric MARV minigenomes mimicking the ZEBOV-specific RNA secondary structure were neither transcribed nor replicated. Mapping of the MARV genomic replication promoter revealed that the region homologous to the sequence involved in formation of the regulatory ZEBOV RNA structure is part of the MARV promoter. The MARV promoter is contained within the first 70 nucleotides of the genome and consists of two elements separated by a spacer region, comprising the TSS of the first gene. Mutations within the spacer abolished transcription activity and led to increased replication, indicating competitive transcription and replication initiation. The second promoter element is located within the nontranslated region of the first gene and consists of a stretch of three UN(5) hexamers. Recombinant full-length MARV clones, in which the three conserved U residues were substituted, could not be rescued, underlining the importance of the UN(5) hexamers for replication activity. Our data suggest that differences in the structure of the genomic replication promoters might account for the different transcription strategies of Marburg and Ebola viruses.
authors.affiliation: Institute of Virology, Philipps University Marburg, Marburg, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Enterlein
authors.forename: Kristina M
authors.initials: KM
authors.lastname: Schmidt
authors.forename: Michael
authors.initials: M
authors.lastname: Schümann
authors.forename: Dominik
authors.initials: D
authors.lastname: Conrad
authors.forename: Verena
authors.initials: V
authors.lastname: Krähling
authors.forename: Judith
authors.initials: J
authors.lastname: Olejnik
authors.forename: Elke
authors.initials: E
authors.lastname: Mühlberger
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D029043:Ebolavirus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral / Q000235:genetics; D006801:Humans; D029024:Marburgvirus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D050506:Mutant Chimeric Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D022661:RNA, Untranslated / Q000235:genetics*; D016415:Sequence Alignment; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation; D014779:Virus Replication
chemical_list: D050506:Mutant Chimeric Proteins; D022661:RNA, Untranslated