Figure 5

Absence of the decay mutants partially enhances Sus1 binding to TREX2 and Mex67. Sus1 association to TREX2 is enhanced in pat1Δ and lsm1Δ. Sus1-TAP was affinity-purified from wild-type (wt), pat1Δ and lsm1Δ strains. The enriched calmoduline eluates from all purifications were analysed by SDS 4-12% gradient polyacrylamide gel electrophoresis and proteins stained with Coomassie. Sus1, Tra1, Spt7 and Sac3 bands were verified by mass spectrometry. (B) Sus1 association to Mex67 is increased in the absence of PAT1 or LSM1. Enrichment of Mex67 in the calmoduline eluates from the respectives deletion strains is demonstrated by western blotting using anti-Mex67 antibodies, whereas no differences were observed for Taf6. (C) Sac3, Mex67 and Sus1 expression levels were confirmed by detecting similar loading from inputs by immunoblot.