Figure 5.

Analysis of the Interactions between the Exosome, the TRAMP, and Nrd1p

(A) Pull-down experiments as in Figure 3B, using Trf4-FLAG as a bait and recombinant Rrp6-Halo, or His-tagged Air2 and Mtr4. The fraction of extract and immunoprecipitated material that is loaded on the gel is indicated.

(B) Western blot analysis of factors associated with the core exosome. Rrp41p-TAP eluates purified from wild-type, trf4Δ, or Δrrp6 cells (IPs) were probed with anti-Rrp6 (1:1,000 dilution) and anti-Trf4 antibodies, respectively.

(C) Pull-down experiment using recombinant Rrp6-Halo as bait and His-tagged Nrd1 variants as indicated. An asterisk indicates a proteolytic fragment of rNrd1 that lacks most of the CID domain.

(D) Schematic summarizing the protein-protein interactions identified in this work. A thinner arrow is used to indicate that the interaction between Rrp6p and Nrd1p cannot be detected in vivo.