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. 2010 Oct 22;22(10):3474–3488. doi: 10.1105/tpc.110.075861

Figure 1.

Figure 1.

Characterization of PUB1 as a Protein Interactor of the LYK3 Intracellular Region.

(A) Domain organization of the PUB1 protein. The U-box domain (amino acids 293 to 364) is located between the UND domain at the N terminus and the ARMADILLO repeats near the C terminus.

(B) PUB1 interacts with LYK3 through the ARM repeats in yeast. Yeast cells expressing either LRRII.1, BRI1, LYK2, LYK4, NFP, or LYK3 intracellular regions cloned in the pBD bait vector were cotransformed with prey constructs cloned in the pGAD vector. The negative control prey consists of the pGAD vector. A full-size PUB1 prey fusion protein was tested for interaction with all bait fusion proteins. Three deletion variants, PUB1ΔUND (1), PUB1ΔUNDΔU-Box (2), and PUB1ΔARM (3), were tested for interaction with the LYK3 bait. Cotransformation of yeast cells with bait and prey vectors is reported by yeast growth on nonselective (NS) medium lacking Trp and Leu. Positive interactions are reported by yeast growth on selective (S) medium lacking Trp, Leu, and His and supplemented with 5 mM 3-amino-triazol. Yeast growth is in black. nd, not determined.