Date of Award
Chemistry and Biochemistry
Erich G. Chapman, Ph.D.
Decay-resistant, s. cerevisiae, XRN1-resistant, xrRNAs, Yeast
Flaviviruses are positive-strand single-stranded RNA viruses that are known to form pseudo-knot RNA structures that halt the progression of 5’→3’ exonuclease Xrn1. We show that these viral Xrn1-resistant structures (xrRNAs) can be used to protect specific homologously-expressed messenger RNAs from 5’→3’ degradation. We investigated the effects of addition of xrRNAs, artificially-installed into the intergenic region of bicistronic mRNA reporters, in the observed levels of protein expression in yeast. The reporters also contain an internal ribosome entry site from the cricket paralysis virus (CrPV IRES) to allow for cap-independent translation of the decay-protected gene, LacZ, encoding the enzyme β-galactosidase. Through the use of primer extension, βgalactosidase assay, and western blots, the results indicate that the partially-decayed RNAs are successfully translated, and that addition of xrRNAs results in a 30-50 fold increase in measured enzymatic activity and an accumulation of decay-resistant transcripts in the cell.
Franklin, Ana Luisa, "Translation of Partially-Decayed Messenger RNAs in Yeast" (2019). Electronic Theses and Dissertations. 1576.
Received from ProQuest
Ana Luisa Franklin
Biochemistry, Molecular biology, Chemistry