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Mutational Analysis of the Rift Valley Fever Virus Glycoprotein Precursor Proteins for Gn Protein Expression
- Source :
- Viruses; Volume 8; Issue 6; Pages: 151, Viruses, Viruses, Vol 8, Iss 6, p 151 (2016)
- Publication Year :
- 2016
- Publisher :
- MDPI AG, 2016.
-
Abstract
- The Rift Valley fever virus (RVFV) M-segment encodes the 78 kD, NSm, Gn, and Gc proteins. The 1st AUG generates the 78 kD-Gc precursor, the 2nd AUG generates the NSm-Gn-Gc precursor, and the 3rd AUG makes the NSm’-Gn-Gc precursor. To understand biological changes due to abolishment of the precursors, we quantitatively measured Gn secretion using a reporter assay, in which a Gaussia luciferase (gLuc) protein is fused to the RVFV M-segment pre-Gn region. Using the reporter assay, the relative expression of Gn/gLuc fusion proteins was analyzed among various AUG mutants. The reporter assay showed efficient secretion of Gn/gLuc protein from the precursor made from the 2nd AUG, while the removal of the untranslated region upstream of the 2nd AUG (AUG2-M) increased the secretion of the Gn/gLuc protein. Subsequently, recombinant MP-12 strains encoding mutations in the pre-Gn region were rescued, and virological phenotypes were characterized. Recombinant MP-12 encoding the AUG2-M mutation replicated slightly less efficiently than the control, indicating that viral replication is further influenced by the biological processes occurring after Gn expression, rather than the Gn abundance. This study showed that, not only the abolishment of AUG, but also the truncation of viral UTR, affects the expression of Gn protein by the RVFV M-segment.
- Subjects :
- 0301 basic medicine
Untranslated region
Recombinant Fusion Proteins
precursor
viruses
DNA Mutational Analysis
030106 microbiology
lcsh:QR1-502
Codon, Initiator
M-segment
Biology
Virus Replication
lcsh:Microbiology
Article
Cell Line
law.invention
Viral Proteins
reverse genetics
03 medical and health sciences
Gaussia
expression strategy
Genes, Reporter
law
Gn
Virology
Animals
Humans
Luciferase
Protein Precursors
Luciferases
Glycoproteins
78 kD
chemistry.chemical_classification
Reporter gene
Rift Valley fever virus
NSm
reporter assay
Gene Expression Profiling
biology.organism_classification
Fusion protein
Molecular biology
030104 developmental biology
Infectious Diseases
chemistry
Viral replication
Recombinant DNA
Glycoprotein
Subjects
Details
- ISSN :
- 19994915
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- Viruses
- Accession number :
- edsair.doi.dedup.....aa76f8028ac891005f8bd22eb2a8ba2b
- Full Text :
- https://doi.org/10.3390/v8060151