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. 2011 Mar 18;23(3):973–983. doi: 10.1105/tpc.110.081653

Figure 1.

Figure 1.

BPEp and ARF8 Proteins Interact in Yeast.

(A) Interaction between BPEp and ARF8 in yeast two-hybrid assays. Yeast cotransformed with pAS2.1-BPEp (BD-BPEp) as bait and pAD-GAL4-2.1-ARF8 (AD-ARF8) as prey or with plasmid vectors without inserts for control of self-activation was diluted three times, and drops were deposited on selective medium A−W−L−and H− (AWLH).

(B) Schematic representation of the BPEp and the ARF8 proteins. The GRSLD motif in SDBPEp that shares sequence similarity with motif III of ARF8 C-terminal domain (CTDARF8) is shown. Conserved amino acids are shown in bold. Key amino acids in the GRSLD motif of SDBPEp that were mutagenized are indicated with asterisks. DBD, DNA binding domain.

(C) Interactions between ARF8 and SDBPEp wild type or mutated at amino acids Arg-250, Asp-253, or Phe-260 were analyzed using β-galactosidase assays in yeast. Mutations of the key amino acids in the GRSLD motif interfere with BPEp and ARF8 interaction. Values are means of three biological replicates. Error bars indicate sd.