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Yeast cells carrying a Sln1-Ypd1 mutant pair with moderate increases in kon and koff displayed a lower threshold of HOG pathway activation than wild-type cells.
This result documents equal intrinsic resistance to the antibody of each mutant of a matched pair, confirming previous biochemical, structural, and genetic studies, which indicated that the epitopes of each mutant pair were indistinguishable regarding reactivity with the monoclonal antibody.
Time-lapse videomicroscopy confirmed that expression of the MKK1/MKK2 mutant pair, which cannot be altered by LT and maintains its ability to signal to MKK1,2 targets including Erk1,2, restored reorientation of the MTOC, leading edge formation and motility (Figure 5C, Movie S9, S10, S11 and S12).
After estimation of w, the interaction class of a mutant pair (a,b) was determined using specific scoring functions S, which transform the fitness matrix into a score matrix <img src="http://journals.plos.org/plosone/article/asset?id=info?doi/10.1371/journal.pone.0011611.e001.PNG" class= inline-graphic"/>.
This concept was supported by a positive relationship between cagA status and Lewis phenotype in clinical H. pylori isolates, as well as in a wildtype and isogenic cagA knockout mutant pair [27], [46] Nevertheless, this correlation is not always obvious, but appears to vary between human populations [46], [55].
Second, correlation between gene expression profiles is calculated for each mutant pair.
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We created mutant pairs of the Sln1-Ypd1 complex interface that caused major compensating changes in the association (kon) and dissociation (koff) rate constants (kinetic perturbations) but only moderate changes in the overall complex affinity (Kd).
Similar results were obtained with at least three independent sets of control and mutant pairs.
Quantitative data were obtained from two independent control littermate and mutant pairs.
In the absence of an established set of known non-interacting mutant pairs, similar to that available for physically non-interacting protein pairs [53], we used here simply the complement of each category as the reference sets of neutral pairs.
The large number of mutant pairs screened, together with a relatively high experimental variability, can make it difficult to extract subtle interaction patterns from the background variability without a sound analytical framework and efficient algorithms [38].
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