Adaptive Processes Change as Multiple Functions Evolve.

Epistasis influences the gene-environment interactions that shape bacterial fitness through antibiotic exposure, which can ultimately affect the availability of certain resistance phenotypes to bacteria. The substitutions present within bla TEM-50 confer both cephalosporin and β-lactamase inhibitor resistance. We wanted to compare the evolution of bla TEM-50 with that of another variant, bla TEM-85, which differs in that bla TEM-85 contains only substitutions that contribute to cephalosporin resistance. Differences between the landscapes and epistatic interactions of these TEM variants are important for understanding their separate evolutionary responses to antibiotics. We hypothesized the substitutions within bla TEM-50 would result in more epistatic interactions than for bla TEM-85 As expected, we found more epistatic interactions between the substitutions present in bla TEM-50 than in bla TEM-85 Our results suggest that selection from many cephalosporins is required to achieve the full potential resistance to cephalosporins but that a single β-lactam and inhibitor combination will drive evolution of the full potential resistance phenotype. Surprisingly, we also found significantly positive increases in growth rates as antibiotic concentration increased for some of the strains expressing bla TEM-85 precursor genotypes but not the bla TEM-50 variants. This result further suggests that additive interactions more effectively optimize phenotypes than epistatic interactions, which means that exposure to numerous cephalosporins actually increases the ability of a TEM enzyme to confer resistance to any single cephalosporin.