Fig. 1

Characterization of the interaction between NSMCE4A and GPS1 by yeast-two-hybrid experiments. (a) Schematic of each GPS1 cDNA prey construct used in the yeast-two hybrid experiments assessing their binding to NSMCE4A. Full length GPS1 is 526 amino acids (aa) long. The schematics include two conserved domains, the RPN7 homology box (PF10602) and the proteasome component (PCI) domain (PS50250). Strength of interaction between each GPS1 prey and NSMCE4A bait is summarized on the right of each prey diagram. (b) Yeast two hybrids grown on a series of selection media to assess interaction between full length NSMCE4A bait and an empty prey vector (negative control), truncated GPS1 prey that cover the N-terminal region (GPS1, 1–288 aa), and C-terminal region (GPS1, 257–526 aa). NSMCE4A bait and GPS1 prey constructs were tested in parallel with a positive bait and prey control (see materials and methods). The media -Leu-Trp does not contain any selection for the interaction. The interaction was tested via the expression from the cassettes encoding Aureobasidin A resistance (AurA), Adenine (−Ade) or Histidine (−His) synthetases. (c) Yeast two hybrid interaction results between full length NSMCE4A bait and specified GPS1 constructs (GPS1, 1–526 aa; GPS1, 257–526 aa; GPS1, 77–526 aa). NSMCE4A bait and GPS1 prey constructs were tested in parallel with a positive and negative bait and prey controls (see materials and methods)