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Three of the four nucleocapsid proteins of Marburg virus, NP, VP35, and L, are sufficient to mediate replication and transcription of Marburg virus-specific monocistronic minigenomes
- Source :
- Journal of virology. 72(11)
- Publication Year :
- 1998
-
Abstract
- This paper describes the first reconstituted replication system established for a member of the Filoviridae, Marburg virus (MBGV). MBGV minigenomes containing the leader and trailer regions of the MBGV genome and the chloramphenicol acetyltransferase (CAT) gene were constructed. In MBGV-infected cells, these minigenomes were replicated and encapsidated and could be passaged. Unlike most other members of the orderMononegavirales, filoviruses possess four proteins presumed to be components of the nucleocapsid (NP, VP35, VP30, and L). To determine the protein requirements for replication and transcription, a reverse genetic system was established for MBGV based on the vaccinia virus T7 expression system. Northern blot analysis of viral RNA revealed that three nucleocapsid proteins (NP, VP35, and L) were essential and sufficient for transcription as well as replication and encapsidation. These data indicate that VP35, rather than VP30, is the functional homologue of rhabdo- and paramyxovirus P proteins. The reconstituted replication system was profoundly affected by the NP-to-VP35 expression ratio. To investigate whether CAT gene expression was achieved entirely by mRNA or in part by full-length plus-strand minigenomes, a copy-back minireplicon containing the CAT gene but lacking MBGV-specific transcriptional start sites was employed in the artificial replication system. This construct was replicated without accompanying CAT activity. It was concluded that the CAT activity reflected MBGV-specific transcription and not replication.
- Subjects :
- Chloramphenicol O-Acetyltransferase
Transcription, Genetic
Immunology
Filoviridae
Chick Embryo
Genome, Viral
Virus Replication
Microbiology
Virus
Chloramphenicol acetyltransferase
Marburg virus
Viral Proteins
Transcription (biology)
Virology
Chlorocebus aethiops
Animal Viruses
Animals
Humans
Viral Regulatory and Accessory Proteins
Replicon
Gene
Vero Cells
Cells, Cultured
Genetics
biology
Nucleocapsid Proteins
biology.organism_classification
Viral replication
Genes
Marburgvirus
Insect Science
RNA, Viral
HeLa Cells
Subjects
Details
- ISSN :
- 0022538X
- Volume :
- 72
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- Journal of virology
- Accession number :
- edsair.doi.dedup.....fe2fc3ac6f2761a617d1f0f951f13038