Figure 5. Nascent transcripts can be oligo-adenylated by the TRAMP complex.

(A) Adenylated (A_n ≥ 3) Mtr4 CRAC reads (blue) superimposed on RNAPI (Rpa190-HTP) 3′ end reads (green). Analyses were performed in rrp44-exo strains, which lack a major exonuclease activity of the exosome complex. Data are represented as median (n = 6) for Rpa190-HTP and mean for Mtr4-HTP rrp44-exo (n = 2).

(B) Metaplot of adenylated (A_n ≥ 3) Mtr4 CRAC reads (blue) superimposed on RNAPI (Rpa190-HTP) 3′ end reads (green).

(C and D) RNAPI (Rpa190-HTP) peaks within the 5′ ETS region (green lines) are reduced and more heterogeneous in trf4-1 trf5Δ and air1Δ air2Δ double-mutant backgrounds (gray lines).

(E and F) RNAPI-associated poly(A) reads in the WT (dark red line) are absent in trf4-1 trf5Δ (and air1Δ air2Δ (gray lines) strains. Note that the lines for the mutants lie along the x axis. For clarity, we display the median of reads in this analysis.

(G) RNAPI (Rpa135-HTP) distribution across the 5′ ETS, comparing WT (black) with rpa12ΔC (gray) strains. Analyses were performed in rrp44-exo strains, which lack a major exonuclease activity of the exosome complex. Data are represented as median (n = 3).

(H) Distribution of Top2-HTP CRAC reads across the 5′ ETS superimposed on RNAPI (Rpa190-HTP) 3′ ends (green). Data are represented as median (Top2-HTP, n = 5; Rpa190-HTP, n = 6).

(I) Metaplot of 3′ ends of adenylated (A_n ≥ 3) Top2 reads (blue) across the 5′ ETS superimposed on RNAPI (Rpa190-HTP) 3′ ends (green).