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Updated: Aug 6, 2026

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
Published on: December 29, 2015
Saccharomyces cerevisiae killer virus transcripts contain template-coded polyadenylate tracts
Abstract:
The M double-stranded RNA component of type 1 killer strains of the yeast Saccharomyces cerevisiae contains an internal 200-base pair adenine- and uracil-rich region. The plus strands of this viral genomic RNA contain an internal adenine-rich region which allows these strands to bind to polyuridylate-Sepharose as tightly as do polyadenylated RNAs with 3'-terminal polyadenylated tracts of 70 to 100 residues. Internal template coding of an adenine-rich tract in positive polarity in vivo and in vitro transcripts of M double-stranded RNA may serve as an alternate method of transcript polyadenylation. The 3'-terminal residue of the in vitro m transcript is a non-template-encoded purine residue. The 5' terminus of this transcript is involved in a stem-and-loop structure which includes an AUG initiation codon, along with potential 18S and 5.8S rRNA binding sites. Except for the 3'-terminal residue, transcription in in vitro shows complete fidelity.
Insights
Killer yeast M double-stranded RNA has an adenine-rich region that mimics polyadenylation, aiding transcript stability and function. This internal RNA feature offers an alternative to traditional 3'-end polyadenylation for viral gene expression.
Area of Science:
- Molecular Biology
- Virology
- Yeast Genetics
Background:
- Type 1 killer strains of Saccharomyces cerevisiae possess M double-stranded RNA (dsRNA).
- This viral dsRNA contains a unique internal adenine- and uracil-rich region.
Purpose of the Study:
- To investigate the function of the internal adenine-rich region in M dsRNA.
- To determine if this region serves as an alternative to 3'-terminal polyadenylation.
Main Methods:
- Analysis of M dsRNA plus-strand binding to polyuridylate-Sepharose.
- In vitro transcription of M dsRNA to analyze transcript structure and fidelity.
Main Results:
- The internal adenine-rich region binds polyuridylate-Sepharose similarly to polyadenylated RNAs.
- In vitro transcripts exhibit a non-template-encoded purine at the 3' terminus.
- The 5' terminus forms a stem-loop structure with an AUG initiation codon and rRNA binding sites.
- In vitro transcription demonstrates high fidelity, except for the 3' terminus.
Conclusions:
- The internal adenine-rich tract in M dsRNA can function as an alternative to 3'-terminal polyadenylation.
- This mechanism may enhance viral transcript stability and translation efficiency in yeast.
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