Figure 4.
Ribosome Traffic Jams at Ribosome Pause Sites.
(A) Measured monosome occupancy from −120 nt to +120 nt around the six leucine codons during leucine starvation. The monosome occupancy was averaged across all occurrences of each codon in the transcriptome.
(B) Measured disome occupancy from −120 nt to +120 nt around the six leucine codons during leucine starvation. The disome occupancy was averaged across all occurrences of each codon in the transcriptome.
(C) Nuclease footprinting assay for detecting ribosome traffic jams on yfp reporter mRNAs. The blue vertical bar along the first variant indicates the location of the CTG200>CTA substitution. Northern blotting was performed using a 32P-labeled antisense RNA complementary to the 300 nt mRNA region from −250 nt to +50 nt of the CTG200>CTA substitution.
(D) Upper panel: Northern blot of nuclease-digested polysomes for the three yfp variants; Lower panel: Polyacrylamide gel corresponding to the Northern blot. Numbers above individual lanes correspond to the three yfp variants in (C). The size markers on the left of the Northern blot were inferred by aligning it to the polyacrylamide gel image. The arrows at 30 nt, 60 nt and 90 nt indicate the approximate locations of monosomes, disomes and trisomes respectively.
See also Figure S3.