An uncertainty law for microbial evolution.

Medical practice would benefit from a thorough understanding of constraints and uncertainty in microbial evolution. Higher order epistasis refers to unexpected effects of multiple mutations even if both single mutations and pairwise effects have been accounted for. Recent studies show that higher order epistasis is abundant in nature, for bacteria as well as higher organisms. However, the importance of higher order effects has been debated. It has been suggested that such effects cannot be interpreted, and should not be considered. Here, we show conclusively that higher order epistasis changes the adaptive prospects for a population. The conclusion is based on an exhaustive search of 193,270,310 hyper-cube graphs and applications of graph theory. Our results are more precise, yet more universal, than related research since they depend on mathematical theory, rather than sampling or simulations. Moreover, the uncertainty we establish for microbial evolution, due to higher order epistasis is not sensitive for detailed model assumptions, such as the baseline being additive or log-additive fitness.