Lsm1-7-Pat1 complex purified from lsm1-14 mutant is weaker than the wild-type complex in binding to the UTR sequences of the BMV RNAs. Uniformly labeled in vitro transcripts carrying different parts (as indicated in the figure) of BMV RNA1 (panel A), RNA2 (panel B), and RNA3 (panel C) were incubated with increasing concentrations of the Lsm1-7-Pat1 complex purified from wild type (WT) or lsm1-14 cells (lanes spanned by black triangles on top) or just BSA as indicated in the figure. The RNA bound to the Lsm1-7-Pat1 complex was then pulled down and visualized by denaturing gel run and phosphorimaging as described in Materials and Methods. Untreated RNA (20% of the input amount) run alongside the bound RNA is also shown. Plot of the percentage of RNA bound (quantitated using the phosphorimager) versus the concentration of the complex used is also shown in each panel.
