Recombinant measles viruses defective for RNA editing and V protein synthesis are viable in cultured cells
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Published on: October 30, 2016
H Schneider1, K Kaelin, M A Billeter
1Institut für Molekularbiologie Abteilung I, Universität Zürich, Hönggerberg, Switzerland.
Abstract:
The measles virus (MV) phosphoprotein (P) gene encodes three proteins, P, C, and V. The V protein is synthesized by pseudo-templated transcription, also designated as RNA editing: during P gene transcription one G residue is inserted at a defined position in about 50% of the mRNAs. To study the importance of sequence elements for the nontemplated G insertion, we generated recombinant MVs in which six different mutations were introduced within the region where editing occurs (3' UUUUUCCC, template strand). These viruses were then analyzed for their ability to edit their P mRNA and to produce V protein. Single U to C changes within the U stretch abolished editing. Extending the template by three C residues at the site of G insertion resulted in a less precise editing phenotype and overproduction of V. None of these mutants were impaired in their multiplication behavior when analyzed in cultured cells. However, the syncytia of a recombinant MV overproducing V protein were in general smaller and lysed 1 to 2 days later than usual.
Insights
Measles virus (MV) RNA editing is crucial for V protein synthesis. Specific sequence elements in the phosphoprotein (P) gene control G-nucleotide insertion, impacting V protein levels and cell syncytia formation.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- The measles virus (MV) phosphoprotein (P) gene encodes P, C, and V proteins.
- V protein synthesis involves a unique RNA editing process: non-templated G insertion into mRNA.
- This editing is essential for V protein production.
Purpose of the Study:
- To investigate the sequence elements critical for non-templated G insertion during MV P gene transcription.
- To understand how mutations in the editing site affect V protein synthesis and viral behavior.
Main Methods:
- Generation of recombinant MVs with six distinct mutations in the P gene's editing region (3' UUUUUCCC).
- Analysis of P mRNA editing efficiency and V protein production in mutant viruses.
- Assessment of viral multiplication in cell culture and examination of syncytia formation.
Main Results:
- Single U to C mutations within the U stretch completely abolished RNA editing.
- Extending the template with three C residues led to imprecise editing and V protein overproduction.
- Mutant viruses showed no impairment in multiplication in cell culture.
Conclusions:
- Specific sequence elements within the MV P gene are essential for accurate RNA editing and V protein synthesis.
- Altered V protein levels, due to impaired editing, can affect measles virus-induced syncytia characteristics.
- These findings provide insights into the molecular mechanisms of MV RNA editing and its functional consequences.
