Overview of experimental design. (A) We replaced the wild-type
allele of the RPO21 (also known as RPB1) gene
with the temperature-sensitive rpb1-1 allele in both the
BY4716 (BY) and the RM11-1a (RM) strains of S. cerevisiae
(Nonet et al. 1987). We mated these
two haploid strains to produce a diploid hybrid and grew the diploid to mid-log
phase at the permissive temperature of 24°C. We rapidly shifted the
temperature of the culture to 37°C, halting transcription. Immediately
following the temperature shift, and at 6, 12, 18, 24, and 42 min after the
shift, we isolated mRNA and performed RNA-seq. (B) By
quantifying the relative levels of the BY and RM alleles for each gene, we
estimated pBY, the proportion of
transcripts derived from BY, at each time point. Under the null hypothesis
(H0; dashed line) of no allele-specific differences in mRNA decay
rates, pBY remains constant. Under
the alternative hypothesis (HA; solid line) of allelic differences
in mRNA decay, we expect pBY to
change as a function of time. For the gene represented by the solid line in the
example pictured, pBY decreases
significantly over time, indicating that the BY allele is decaying more quickly
than the RM allele of this gene.