Figure 2

Yeast two-hybrid array screens and vectors. Shown are two Y2H screens with four different vector combinations. Each interaction is represented by two colonies to ensure reproducibility. (A) Lambda bait protein A (DNA packaging protein) was fused to an N-terminal DNA-binding domain ("DBD", in pGBKT7g) and was tested against prey constructs in both N- and C-terminal configurations (activation domains in pGADT7g, and pGADCg). (B) The C-terminal DBD fusion (in pGBKCg) as tested against prey constructs in both N- and C-terminal configurations (in pGADT7g, and pGADCg). The interactions of C-terminal preys are labeled with an asterisk (*), all remaining interactions use N-terminal fusions. All the interactions obtained from the array screening were subjected to Y2H retests: we were able to retest all the interactions shown in Figure 2 except A-Ea47, which has thus been removed from the final interaction list. Technical details of the screening procedure have been described in [8, 10]. (C) Interaction quality assesment. Using the experimental derived false positive rate from [9] and Bayes theorem, we estimated the probability of an interaction to be true. This estimate depends on the vector system, being highest (83%) for pDEST22/32, and lowest (40%) for pGBKCg/pGADT7g. (D) Detection of known PPIs with different vector systems. Known PPIs are enriched in the subset of PPIs detected by > = 2 vector systems compared to PPIs detected by 1 vector combination.