Sequence variability in viral genome non-coding regions likely contribute to observed differences in viral replication amongst MARV strains

The Marburg viruses Musoke (MARV-Mus) and Angola (MARV-Ang) have highly similar genomic sequences. Analysis of viral replication using various assays consistently identified MARV-Ang as the faster replicating virus. Non-coding genomic regions of negative sense RNA viruses are known to play a role in viral gene expression. A comparison of the six non-coding regions using bicistronic minigenomes revealed that the first two non-coding regions (NP/VP35 and VP35/VP40) differed significantly in their transcriptional regulation. Deletion mutation analysis of the MARV-Mus NP/VP35 region further revealed that the MARV polymerase (L) is able to initiate production of the downstream gene without the presence of highly conserved regulatory signals. Bicistronic minigenome assays also identified the VP30 mRNA 5′ untranslated region as an rZAP-targeted RNA motif. Overall, our studies indicate that the high variation of MARV non-coding regions may play a significant role in observed differences in transcription and/or replication.

► Differences in the replication rates of MARV strains were observed.
► MARV-Ang consistently produced relatively greater amount of viral mRNA and protein.
► Two non-coding viral genomic regions affect expression of reporter genes at significantly different rates.
► Additional MARV UTR rZAP targets identified using bicistronic mini-genome system.
► MARV-Mus NP/VP35 RR able to initiate transcription of downstream gene in the absence of highly conserved stop/start signals.