A Novel Role for the TIR Domain in Association with Pathogen-Derived Elicitors
Figure 8
N's TIR Domain Is Sufficient for Association with p50-U1
(A) Co-immunoprecipitation of gN-TIR-TAP and p50-U1-Cerulean. N's TIR domain was expressed under the control of N's endogenous 5′ and 3′ regulatory regions. Extracts from tissue co-expressing N(TIR)-TAP (top panel, lanes 1 and 2) and p50-U1-Cerulean (middle panel, lane 1) or p50-U1-Ob-Cerulean (middle panel, lane 2) were incubated with anti-GFP antibodies. Immunoprecipitated complexes were separated by SDS-PAGE and probed with anti-MYC antibodies. N(TIR)-TAP was pulled down with p50-U1-Cerulean (bottom panel, lane 1), but not with p50-U1-Ob-Cerulean (bottom panel, lane 2). Lane M is the size marker, and protein sizes are shown in kDa.
(B) BiFC between N(TIR)-YN and p50-U1-YC. N(TIR)-YN exhibits BiFC with p50-U1-YC (column 1), but not with p50-U1-Ob-YC (column 2). The TIR domains of two related R proteins, BS4 and RPP5, were tested for their ability to associate in vivo with p50-U1-YC. BS4(TIR)-YC and RPP5(TIR)-YC were co-expressed with p50-U1-YC, but were unable to exhibit BiFC (columns 3 and 4, respectively). Scale bar represents 20 μm.