A novel test for selection on cis-regulatory elements reveals positive and negative selection acting on mammalian transcriptional enhancers

TL;DR

This paper introduces a new method to detect positive and negative selection on mammalian transcriptional enhancers by analyzing mutation effects and evolutionary patterns, revealing diverse selection modes across different enhancers.

Contribution

The study presents a novel approach using saturation mutagenesis data and phylogenetic analysis to measure selection on cis-regulatory elements, advancing understanding of their evolutionary dynamics.

Findings

All enhancers studied show negative selection against down-regulating mutations.

One enhancer exhibits positive selection for up-regulating mutations.

Other enhancers show no detectable selection on up-regulating mutations.

Abstract

Measuring natural selection on genomic elements involved in the cis-regulation of gene expression -- such as transcriptional enhancers and promoters -- is critical for understanding the evolution of genomes, yet it remains a major challenge. Many studies have attempted to detect positive or negative selection in these noncoding elements by searching for those with the fastest or slowest rates of evolution, but this can be problematic. Here we introduce a new approach to this issue, and demonstrate its utility on three mammalian transcriptional enhancers. Using results from saturation mutagenesis studies of these enhancers, we classified all possible point mutations as up-regulating, down-regulating, or silent, and determined which of these mutations have occurred on each branch of a phylogeny. Applying a framework analogous to Ka/Ks in protein-coding genes, we measured the strength of selection on up-regulating and down-regulating mutations, in specific branches as well as entire phylogenies. We discovered distinct modes of selection acting on different enhancers: while all three have experienced negative selection against down-regulating mutations, the selection pressures on up-regulating mutations vary. In one case we detected positive selection for up-regulation, while the other two had no detectable selection on up-regulating mutations. Our methodology is applicable to the growing number of saturation mutagenesis data sets, and provides a detailed picture of the mode and strength of natural selection acting on cis-regulatory elements.