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. 2012 Jan 30;158(4):1628–1642. doi: 10.1104/pp.111.191965

Figure 1.

Figure 1.

Interaction assays between GDU family members and the E3 ubiquitin ligases of the LOG2 family. A, yeast two-hybrid interaction of the cytosolic domain of the GDU proteins (cGDU) with LOG2 and LULs. The panels show swapping of inserts between the bait (pGBT9) and prey (pACT2) plasmids. Yeast coexpressing the protein pairs were grown for 4 d on medium selecting for protein interaction, lacking Leu, Trp, adenine, and His. All yeast grew on a medium lacking Leu and Trp, selecting for the plasmids only (data not shown). B, GST pull-down (PD) assay of flag-cGDU1 or flag-cGDU1G100R with GST-LOG2, GST-LUL1, or GST alone. Top, pulled-down samples; middle, cGDU input; bottom, GST protein input. Arrowheads indicate GST (approximately 27 kD) or full-length GST-tagged LOG2 and LUL1. C, Coimmunoprecipitation (IP) assay after the expression of GDU1 or GDU1G100R and mLOG2 or mLOG2R12K in transiently infiltrated N. benthamiana leaves. Top, Myc coimmunoprecipitation samples probed with α-HA (H); middle, Myc coimmunoprecipitation samples probed with α-Myc (M); bottom, HA-protein input. Asterisks and diamonds indicate LOG2 and GDU1 proteins, respectively. Numbers on the right in B and C indicate molecular mass in kD. ORF, Open reading frame; WB, western blot.