Recombinant measles viruses defective for RNA editing and V protein synthesis are viable in cultured cells.
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.
Schneider
H
H
Institut für Molekularbiologie Abteilung I, Universität Zürich, Hönggerberg, Switzerland.
Kaelin
K
K
Billeter
M A
MA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D003673:Defective Viruses / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008459:Measles virus / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017393:RNA Editing; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*; D014779:Virus Replication
D017931:DNA Primers; D010750:Phosphoproteins; C069181:V protein, measles virus; D014764:Viral Proteins