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. 2005 Jan;137(1):83–93. doi: 10.1104/pp.104.052654

Figure 5.

Figure 5.

Mutagenesis of conserved residues in AtBPM1 affects assembly with AtCUL3 and AtBPM1 proteins. A, Schematic overview of AtBPM1 and locations of mutagenized amino acid residues (underlined and pointed out by arrows). B, In contrast with AtBPM1 and AtBPM1D204N, AtBPM1L189Stop and AtBPM1D204A did not assemble with AtCUL3a and AtBPM1 proteins in Y2H assays. In addition, AtBPM1D328A and AtBPM1Y330A strongly reduced interaction with AtCUL3a and AtBPM1. The introduction of a point mutation at position 332 (AtBPM1L332A) only affected interaction with AtBPM1. C, Results from the Y2H studies were verified for AtBPM1D204A and AtBPM1L189STOP by pull-down assays with in vitro translated AtCUL3a, AtBPM1 and mutated AtBPM1 versions, and bacterially expressed and purified GST, GST∷AtBPM1, GST∷AtBPM1D204A, or AtBPM1L189STOP fusion proteins.