EXPRESSION OF DOUBLE-STRANDED RNA IN EUKARYOTIC CELLS

Project: Research project

Description

The cytoplasmically-inherited killer virus of yeast will be used as a model
to study the expression of the information on a double-stranded RNA genome
in a eukaryotic cell. This virus contains a virion-associated RNA
polymerase activity which catalyzes the transcription in vitro of a
message-polarity single strand of RNA which is released from the virion.
In heterologous eukaryotic translation systems, this encodes a 32,000
dalton protein which is apparently identical to the in vivo precurser of
the low molecular weight killer toxin protein produced by killer yeast
strains which harbor the virus. Transcription and translation in vitro by
homologous yeast systems will be used to study this genetically
well-characterized virus. Transcription by isolation virions purified from
yeast cytoplasm will be studied to elucidate the priming and initiation
mechanism of the viral RNA polymerase. Transcripts and isolated separated
genomic RNA strands will be tested as translational templates, with
attention focused upon structural features required for message activity in
the yeast system. Open reading frames elucidated by sequence analysis of
RNA will be tested in their function by ribosome binding experiments and
sequence determination of the primary translation products.
Post-transcriptional modifications of the messenger RNA including cap
structures and polyadenylation will be tested for their effects on
translation template activity. Thus, the killer virus of yeast will be
used to elucidate the mechanisms controlling transcription and translation
in eukaryotic cells, processes important in regulation of many cellular
functions.
StatusFinished
Effective start/end date7/1/836/30/87

Funding

  • National Institutes of Health

Fingerprint

Double-Stranded RNA
Eukaryotic Cells
Viruses
Yeasts
Virion
RNA
Polyadenylation
Viral RNA
DNA-Directed RNA Polymerases
Ribosomes
Open Reading Frames
Sequence Analysis
Cytoplasm
Proteins
Molecular Weight
Messenger RNA

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)